As a prerequisite for sexual reproduction,gametophyte development is an interesting process involving cell proliferation,differentiation and specialization.Gametogenesis has been extensively explored in model plants,b...As a prerequisite for sexual reproduction,gametophyte development is an interesting process involving cell proliferation,differentiation and specialization.Gametogenesis has been extensively explored in model plants,but the regulatory mechanism of gametophyte development largely remains unknown in Cucurbitaceae species.In present study,we have shown that watermelon ClBBM and ClPLT2,two AP2/ERF transcription factors,participated in both male and female gametophyte development.Clbbm and Clplt2 single mutants resembled wild-type phenotypes in both vegetative and reproductive development.But Clbbm/Clplt2 double mutant showed partial pollen abortion and bore less seeds comparing to WT.Our results indicated that the abnormal pollen grains were caused by premature tapetum degeneration,and reduced seed-set was due to faulty embryo sac development.ClBBM and ClPLT2 were expressed in FG4 embryo sacs,and their transcripts were also detectable in the tapetum and microspore of stage 9 anthers,which was consistent with developmental stages of defective phenotypes observed in double mutant.The expression of genes essential for tapetum development,ClATM1,ClAMS,ClMS1 and ClMS188,was decreased in Clbbm/Clplt2 double mutants.Moreover,the transcriptome analysis indicated that ClBBM and ClPLT2 participated in tapetum and pollen wall development by regulate cell cycle,transmembrane transport,glucan and cellulose metabolic process.Collectively,ClBBM and ClPLT2 were functionally redundant in regulating gametophyte development in watermelon,and their functions differ from their homologous genes in model plant Arabidopsis.展开更多
An interspecific hybrid F1 of Cucumis hystrix Chakr. x Cucumis sativus L. （NC4406） was used to establish the developmental sequence and to characterize the male and female gametophytes at cytological level for furth...An interspecific hybrid F1 of Cucumis hystrix Chakr. x Cucumis sativus L. （NC4406） was used to establish the developmental sequence and to characterize the male and female gametophytes at cytological level for further understanding of the phylogenic relationship and the mechanism of fertility or sterility in the interspecific hybrid F1 The development of male and female gametophytes was studied through meiotic analysis and paraffin section observation technique, respectively. Meanwhile, the fertility level was assessed through hybrid F1 backcrossing to cultivated cucumber 4406. Variable chromosome configurations were observed in the pollen mother cells （PMCs） of hybrid F1 at metaphase Ⅰ , e.g., univalents, bivalents, trivalents, quadravalents, etc. At anaphase Ⅰ and Ⅱ, chromosome lagging and bridges were frequently observed as well, which led to the formation of polyads and only a partial number of microspores could develop into fertile pollen grains （about 23.3%）. Observations of the paraffin sections showed numerous degenerated and abnormal embryo sacs during the development of female gametophytes, and only 40% of the female gametophytes could develop into normal eight-nuclear megaspore. On an average, 22.8 and 6.3 seeds per fruit could be obtained from the reciprocal backcross. The interspecific hybrid F1 of C. hystrix x NC4406 was partially fertile; however, the meiotic behaviors of hybrid F1 showed a high level of intergenomic recombination between C. hystrix and C. sativus chromosomes, which indicated that it plays an important role for introgression of useful traits from C. hystrix into C. sativus.展开更多
Dear Editor, Environmental arsenic (As) contamination of both soil and water systems is a worldwide problem, and As can enter the human food chain through contaminated agricultural products and drinking water and cont...Dear Editor, Environmental arsenic (As) contamination of both soil and water systems is a worldwide problem, and As can enter the human food chain through contaminated agricultural products and drinking water and contribute to the development of lung.展开更多
Gametophytes of Laminaria japonica were cryopreserved in liquid nitrogen using encapsulation-dehydration with two-step cooling method. Gametophytes cultured at 10℃ and under continuous irradiance of 30 μmol m^-2 s^-...Gametophytes of Laminaria japonica were cryopreserved in liquid nitrogen using encapsulation-dehydration with two-step cooling method. Gametophytes cultured at 10℃ and under continuous irradiance of 30 μmol m^-2 s^-1 for 3 weeks were encapsulated in calcium alginate beads. The beads were dehydrated in 0.4 molLl sucrose prepared with seawater for 6 h, desiccated in an incubator set at 10℃ and 70% relative humidity for 4 h, pre-frozen at either -40℃ or -60℃ for 30 min, and stored in liquid nitrogen for 〉24 h. As high as 43% of survival rate was observed when gametophytes were thawed by placing the beads in 40℃ seawater and re-hydrated in 0.05 molL^-1 citrate sodium prepared using 30‰ NaCl 7 d later. More cells of male gametophytes survived the whole procedure in comparison with female gametophytes. The cells of gametophytes surviving the preservation were able to grow asexually and produce morphologically normal sporophytes.展开更多
A newly identified bacterial disease of kelp(Saccharina japonica) gametophytes was found in clone cultures.It is characterized by swollen gametophyte cells in the early period of infection followed by filamentous fadi...A newly identified bacterial disease of kelp(Saccharina japonica) gametophytes was found in clone cultures.It is characterized by swollen gametophyte cells in the early period of infection followed by filamentous fading.An alginolytic marine bacterium referred to as A-1 was isolated from the diseased gametophytes.On the basis of 16S rDNA sequencing and morphological,physiological and biochemical characteristics,the bacterium was identified as a strain of the genus Alteromonas.By testing Koch's postulates,Alteromonas sp.A-1 was further confirmed as the pathogen.The infection process was also investigated using both scanning electron and light microscopy.展开更多
This study on the effects of ultrasonic treatment on female gametophytes of Laminariajaponica showed that:1. Ultrasonic treatment had shortening effect on filaments of female gametophytes. Within certainperiod of time...This study on the effects of ultrasonic treatment on female gametophytes of Laminariajaponica showed that:1. Ultrasonic treatment had shortening effect on filaments of female gametophytes. Within certainperiod of time, the average length of filamentous female gametophytes was shortened.2. Ultrasonic treatment had emptying effect on cells. The number of empty cells increased with timeof treatment. Ultrasonic treatment had harmful effect on cells.3. Ultrasonic treatment could break down cell walls. The combination of frequency of 20 kHz, out-put of 15 W, 40 s and 60 s of treatment was best for this purpose. After ultrasonic treatment, the regner-ation of female gametophytes into sporophytes was effected. Female gametophytes could not recover aftertoo long period of treatment.展开更多
The annual life cycle of the brown seaweed Undariapinnatifida （Harvey） Suringer comprises a macroscopic diploid sporophyte stage and a microscopic haploid gametophyte stage. In 2011, an unusual zoospore-derived mono...The annual life cycle of the brown seaweed Undariapinnatifida （Harvey） Suringer comprises a macroscopic diploid sporophyte stage and a microscopic haploid gametophyte stage. In 2011, an unusual zoospore-derived monoecious gametophyte isolate （designated as line 10-5-3） of U. pinnatifida was observed. To understand this phenomenon, a comprehensive screening of eighty-two previously identified male gametophyte cultures, isolated from three randomly selected cultivars （lines 10, 7, and 5） was performed. Thirty-six of the isolates developed both antheridia and oogonia on the same filamentous fragment in a standard gametogenesis test （SGT： 18℃, 60 umol photons/（m2.s））. Selfing of the monoecious gametophyte or crossing it with a normal male gametophyte both gave rise to morphologically normal sporophytic offspring. However, crossing resulted in a much higher fertilization rate （89.7%）. The hybrid and selfed sporophytic offspring were grown to maturity in flow tanks at an ambient temperature of 10-18℃ over a period of 69 days. Active zoospores were released from both types of mature sporophylls. The majority of these developed into male gametophytes, while 15%-20% developed into the observed monoecious structures on the same filament. Using PCR amplification it was found that all the monoecious gametophyte isolates and the sporophytic offspring resulting from the selfing and crossing lacked the femalelinked microsatellite sequence （a part of the locus Up-AC-2A8, GenBank accession No. AY738602.1）, indicating their male nature. U.pinnatifida is an invasive species in some regions and the implications of the above findings for this species in nature are briefly discussed.展开更多
The objective of this study was to examine the eff ects of temperature and light intensity on growth of female gametophytes of Eisenia bicyclis and Ecklonia cava and responses of these female gametophytes to Fe additi...The objective of this study was to examine the eff ects of temperature and light intensity on growth of female gametophytes of Eisenia bicyclis and Ecklonia cava and responses of these female gametophytes to Fe addition and daylength. Female gametophytes of each species were cultured at four temperatures (10, 15, 20, and 25°C) and under a combination of four light intensities (10, 20, 40, and 80 μmol photons/(m^2 ·s)) with two temperatures (15 and 20°C for E i . bicyclis;20 and 25°C for E c . cava ) to clarify their optimal growth conditions. Growth and maturation of female gametophytes of these two species under a combination of five Fe-EDTA concentrations (0, 1, 2, 4, and 8 μmol/L) and three daylengths (10, 12, and 14 h) were also examined. The growth of E i . bicyclis gametophyte was maximal at approximately 15–20°C, 20 μmol photons/(m^2 ·s), Fe-EDTA concentration of 8 μmol/L and daylengths of 12–14 h. While E c . c ava gametophytes showed optimal growth at approximately 20–25°C, 20 μmol photons/(m^2 ·s), Fe- EDTA concentration of 8 μmol/L and daylength of 14 h. Maturation of gametophytes was enhanced at Fe-EDTA concentration of 4 μmol/L for E i . bicyclis and at 2 μmol/L for E c . c ava . In conclusion, optimal growth temperatures and Fe-EDTA concentrations for maturation of E i . bicyclis and E c . c ava gametophytes were diff erent. Higher optimal growth temperature for E c . c ava gametophytes may contribute to its wider geographical distribution compared to E i . bicyclis which has restricted habitats in Korea. This suggests that addition of Fe ion could be used to recover beds of these two species in barren grounds of Korea.展开更多
Abstract: Inter-simple sequence repeat (ISSR) analysis was used to assess genetic diversity among 10 pairs of male and female Laminaria gametophytes. A total of 58 amplification loci was obtained from 10 selected ISSR...Abstract: Inter-simple sequence repeat (ISSR) analysis was used to assess genetic diversity among 10 pairs of male and female Laminaria gametophytes. A total of 58 amplification loci was obtained from 10 selected ISSR primers, of which 34 revealed polymorphism among the gametophytes. Genetic distances were calculated with the Dice coefficient ranging from 0.006 to 0.223. A dendrogram based on the unweighted pair-group method arithmetic (UPGMA) average showed that most male and female gametophytes of the same species were clustered together and that 10 pairs of gametophytes were divided into four groups. This was generally consistent with the taxonomic categories. The main group consisted of six pairs of gametophytes, which were selected from Laminaria japonica Aresch. by intensive inbreeding through artificial hybridization. One specific marker was cloned, but was not converted successfully into a sequence characterized amplified region (SCAR) marker. Our results demonstrate the feasibility of applying ISSR markers to evaluate Laminaria germplasm diversities.展开更多
Carbonic anhydrase(CA)was considered to be an important component of carbon concentrating mechanism(CCM)of algae.It was an inducible enzyme.Environmental factors,especially dissolved inorganic carbon and pH,were known...Carbonic anhydrase(CA)was considered to be an important component of carbon concentrating mechanism(CCM)of algae.It was an inducible enzyme.Environmental factors,especially dissolved inorganic carbon and pH,were known to affect CA activity.Effects of inorganic carbon(CO_(2)and HCO_(3)^(-))and pH on CA activity of gametophytes of Saccharina japonica were evaluated in this study.Under high-CO_(2)condition(3%CO_(2)),the activity of external CA(C_(Aext))was significantly decreased(P<0.05)from 33.92 REA/g FW to 27.69 REA/g FW.In contrast,the internal CA(CA_(int))and total CA activities were elevated significantly(p<0.01)from 36.83 REA/g FW to 48.80 REA/g FW,and from 70.75 REA/g FW to 76.49 REA/g FW,respectively.Addition of an appropriate concentration of HCO_(3)^(-)to the medium,CA_(int)and total CA activities were promoted significantly,although the activity of C_(Aext)was inhibited significantly(P<0.05).In higher HCO_(3)^(-)concentration(up to 1680 mg/L),the activity of CA_(int)was promoted to 60.81REA/g FW,which was twice that of the control.Higher activities of CA_(int)and total CA were induced at low pH.At pH 6.0,the highest activities of CA_(int)(110.85 REA/g FW)and total CA(128.17 REA/g FW)were induced.However,at pH 9.0,the activities of CA_(int)and total CA were reduced to the lowest of 23.31REA/g FW and 42.19 REA/g FW,respectively.This is the first report about the successful detection of C_(Aext)activity of gametophytes of S.japonica.The results would provide data for the analysis of Ci acquisition and transport mechanism in S.japonica gametophytes.展开更多
The life cycle of seaweed Laminaria japonica involves a generation alternation between diploid sporophyte and haploid gametophte.The expression of foreign genes in sporophte has been proved.In this research,the recomb...The life cycle of seaweed Laminaria japonica involves a generation alternation between diploid sporophyte and haploid gametophte.The expression of foreign genes in sporophte has been proved.In this research,the recombinant expression in gametophyte was investigated by particle bombardment with the rt-PA gene encoding the recombinant human tissue-type plasminogen activator(Reteplase),which is a thrombolytic agent for acute myocardial infarction(AMI).Transgenic gametophytes were selected by their resistance to herbicide phosphiothricin(PPT),and proliferated in an established bubble column photo-bioreactor.According to the results from quantitative ELISA,Southern blotting,and fibrin agarose plate assay(FAPA)for bioactivity,it was showed that the rt-PA gene had been inte-grated into the genome of gametophytes of L.japonica,and the expression product showed the ex-pected bioactivity,implying the proper post-transcript modification in haploid gametophyte.展开更多
Effects of various concentrations of two heavy metals, namely Cd and Cu, on gametophytes of Laminariajaponica Aresch were determined by recording morphological changes of gametophytes, determining pH values and the he...Effects of various concentrations of two heavy metals, namely Cd and Cu, on gametophytes of Laminariajaponica Aresch were determined by recording morphological changes of gametophytes, determining pH values and the heavy metal content of the culture solution, calculating the germination rate of sporophytes, and observing heavy metal （Cd） distribution using a fluorescence microscope. The results showed that heavy metals damaged the gametophytes, and were even lethal, and that the higher the concentration of heavy metal ions, the greater the injury to gametophytes. Gametophytes could not survive in culture solutions containing more than 100 mg/L Cd and 50 mg/L Cu and were only able to survive in culture solution containing a mixture of Cd and Cu up to a concentration of 10 mg/L, which indicates that gametophytes have a higher tolerance to Cd than Cu and that multiple heavy metal ions in solution markedly aggravate the damage to gametophytes compared with individual heavy metal ions. With increases in the concentration of the heavy metal, the burgeoning rate of sporophytes decreased acutely, and solutions containing multiple heavy metal ions caused even more marked harm to sporophytes than solutions containing a single heavy metal ion, because most sporophytes died in mixed solutions. The pH value of the culture medium dropped immediately at the beginning （the first day） of treatment, increased over the following days, and then decreased again. The pH of culture media containing multiple heavy metal ions showed greater variation than media containing a single heavy metal ion, with the extent of the decrease in pH of culture media containing multiple ions being greatest during the last period of the experiment. With increases in the concentration of heavy metals, the capacity of gametophytes to accumulate these ions increased. The blue fluorescent light emitted by the Cd-and Cd-binding protein complex existing in gametophytes in media containing different concentrations of Cd showed clearly the distribution of the ion in gametophytes and the results obtained were consistent with distribution determined using other methods. All results of the present study showed that gametophytes of L. japonica play a remarkable role as heavy metal decontaminators, especially with regard to Cd.展开更多
[Objective] Pollen mother cell miosis and male gametophyte development of pumpkin were observed in this study, to provide some cytological basis for pumpkin anther or microspore culture. [Method] Ehrlich＇s hematoxyli...[Objective] Pollen mother cell miosis and male gametophyte development of pumpkin were observed in this study, to provide some cytological basis for pumpkin anther or microspore culture. [Method] Ehrlich＇s hematoxylin staining-methyl salicylate clearing technique was used for observation and research of the variation of cell structure and chromosomal behavior during pollen mother cell miosis and male gametophyte development of ‘Tianhong＇ pumpkin. [Result] The meiosis in pollen moth- er cells of pumpkin was simultaneous cytokinesis. In the process of nuclear division, nuclear membrane and nucleolus of pumpkin pollen mother cells gradually disappeared in the metaphase I and reappeared in telophase I , phragmoplast formed between the two generated crescent-shaped nuclei without cell wall, the phragmoplast gradually disappeared in the metaphase II and reappeared in telophase II. Phragmoplast spread outward from the center of spindle during the second division was connected with that formed on the central interface of two nuclei during the first division, cell wall of microspores generated from periphery to center. Most of the tetrads contained four sub-cells while a few contained extra small cells. During the period of uniuclete microspore at periphery, the single nucleolus split into 2-3 or more small nucleoli, mature pollen grain was two-celled. Mononucleate pollen cells were mostly appeared in the flower buds with length of 1.0-2.0 cm, which could be used as an important indicator to collect materials for anther or microspore culture. [Conclusion] This study laid the foundation for research of the cytogenetics of pumpkin.展开更多
Abstract Ulva linza is one of the species that causes green tides in the Yellow Sea,China.Due to the diffi culties in chromosomal preparation,the large numbers of chromosomes,and their relatively small sizes,there hav...Abstract Ulva linza is one of the species that causes green tides in the Yellow Sea,China.Due to the diffi culties in chromosomal preparation,the large numbers of chromosomes,and their relatively small sizes,there have been no reported studies on Ulva macroalgae chromosomes.The karyotypes and chromosomes in U.linza were observed after a series of treatments.The chromosomes were pretreated with 0.1%colchicine for 12 h and then mixed with enzymes.The samples were dropped from 30 cm height onto glass slides,which spread out the surface coat.These pretreatments were the optimal chromosomes preparation treatments.The prepared chromosomes were stained with 4',6-diamidino-2-phenylindole(DAPI),which is a fl uorescent probe sensitive and specifi c to DNA.The chromosome number in the haploid male and female gametophytes was n=9,and was 2 n=18 in the diploid sporophytes.The female gametophyte chromosomes were between 0.804 and 2.292μm in size,the male gametophyte chromosomes were between 0.917 and 2.916μm,and the sporophyte chromosomes were between 0.912 and 2.167μm.The relative sizes of the chromosomes were used to analyze the karyotypes of the female and male gametophyte chromosomes.The results provide a solid foundation for the basic technique that can be used to localize molecular markers of Ulva chromosomes.展开更多
Eighteen gametophytes includingL. japonica, L. ochotensisandL. longissima, were verified with random amplified polymorphic DNA (RAPD) technique. Eighteen ten-base primers were chosen from 100 primers selected for fina...Eighteen gametophytes includingL. japonica, L. ochotensisandL. longissima, were verified with random amplified polymorphic DNA (RAPD) technique. Eighteen ten-base primers were chosen from 100 primers selected for final amplification test. Among the total of 205 bands amplified, 181 (88.3%) were polymorphic. The genetic distance among different strains ranged from 0.072 to 0.391. The dendrogram constructed by unweighted pair-group method with arithmetic (UPGMA) method showed that the female and male gametophytes of the same cell lines could be grouped in pairs respectively. It indicated that RAPD analysis could be used not only to distinguish different strains ofLaminaria, but also to distinguish male and female gametophyte within the same cell lines. There is ambiguous systematic relationship if judged merely by the present data. It seems that the use of RAPD marker is limited to elucidation of the phylogenetic relationship among the species ofLaminaria.展开更多
Unicellular gametophytes of Undaria pinnatifida (Harv.) Sur. were isolated in Qingdao, P. R. China in April 1993 and in Tokushima, southem Japan in March 1995. Diferent intraspecific crossings by using unicellular mal...Unicellular gametophytes of Undaria pinnatifida (Harv.) Sur. were isolated in Qingdao, P. R. China in April 1993 and in Tokushima, southem Japan in March 1995. Diferent intraspecific crossings by using unicellular male and female gametophytes were successfully undertaken in Sept. of 1995 in Qingdao.Sporophytes were transplanted to two different locations for open an cultivation. One was at Zhanqiao (ZQ) Bay where the water current was slower than that of another location - Taipingjiao(TPJ). A total of218 adult sporophytes were harvested on January 12 and 18 from TPJ and ZQ repectively. For eacncombination, 10 sporophytes un cultivated. Analysis of the mprpholgical characteristics of adult spprophytes indicated that the longest length between two bases of the serration of pinnate bldes (W2) is a morphological characteristic that can be transferred from the parent plant to the next generations regardless of environmental variations. There was evidence that W2 was apparently determined by sex-linked factors, i.e, by male parental grametophyte.. Sporophytes from certain crossing combinations showed more vigorous growth than those from other crossing combinations. It is therrfore possible to select grametophyte strains which can be used as parental gametophytes for the seeding production of sporophytes with more vigorous growth within shorter cultivation period. The morphology of hybrids from a Qingdao strain and a Tokushima cultivated strain resembled that of both parental plants in frond feaures (wrinkled or smooth) and W2. Sporophyll formation also varied with strains. The fact that adult sporophytes resulting from the same crossing combinations have identical morphological characteristics under the same environmental conditions indicates the possibility of a new way to select strains which are expected to be ideal for commerical production by purposely selecting, propagating, and seeding unicellular gametophytes for sporeing production through freeliving techniques of gametophytes.展开更多
Suitable habitat is vital for the survival and restoration of a species.Understanding the suitable habitat range for lycophytes and ferns is prerequisite for effective species resource conservation and recovery effort...Suitable habitat is vital for the survival and restoration of a species.Understanding the suitable habitat range for lycophytes and ferns is prerequisite for effective species resource conservation and recovery efforts.In this study, we took Athyrium brevifrons as an example, predicted its suitable habitat using a Maxent model with 67 occurrence data and nine environmental variables in Northeast China.The area under the curve(AUC) value of independent test data, as well as the comparison with specimen county areal distribution of A.brevifrons exhibited excellent predictive performance.The type of environmental variables showed that precipitation contributed the most to the distribution prediction, followed by temperature and topography.Percentage contribution and permutation importance both indicated that precipitation of driest quarter(Bio17) was the key factor in determining the natural distribution of A.brevifrons, the reason could be proved by the fern gametophyte biology.The analysis of high habitat suitability areas also showed the habitat preference of A.brevifrons: comparatively more precipitation and less fluctuation in the driest quarter.Changbai Mountains, covering almost all the high and medium habitat suitability areas, provide the best ecological conditions for the survival of A.brevifrons, and should be considered as priority areas for protection and restoration of the wild resource.The potential habitat suitability distribution map could provide a reference for the sustainable development and utilisation of A.brevifrons resource, and Maxent modelling could be valuable for conservation management planning for lycophytes and ferns in Northeast China.展开更多
The cultivation of the first filial generation of monoploid gametophyte clones of different Laminaria species (hybrid Laminaria) is an effective way of utilizing heterozygous vigor (heterosis). A female gametophyte cl...The cultivation of the first filial generation of monoploid gametophyte clones of different Laminaria species (hybrid Laminaria) is an effective way of utilizing heterozygous vigor (heterosis). A female gametophyte clone of L. japonica and a male gametophyte clone of L. longissima were hybridized, generating Dongfang No.2 hybrid Laminaria. The parentage of this hybrid Laminaria was determined using AFLP of total DNA, SNP of the ITS region of ribosomal RNA transcription unit and microsatellite DNA variation at two loci. In addition to 167 AFLP bands shared by Dongfang No.2, L. japonica and L. longissima, Dongfang No.2 hybrid Laminaria shared another 70 and 55 bands with L. japonica and L. longissima, respectively, which were obviously more than 11 bands shared by L. japonica and L. longissima. Dongfang No.2 held both ‘T’ and ‘C’ at position 847 of the ITS region, while ‘T’ at this position was specific for L. japonica and ‘C’ for L. longissima, respectively. Dongfang No.2 also held the microsatellite DNA alleles of two parents together at two microsatellite DNA marker loci. These observations clearly proved that Dongfang No.2 is a true hybrid of L. japonica and L. longissiuma. Unfortunately, the origin of the chloroplast of Dongfang No.2 was not determined based on the variation of RuBisCo spacer. More sequence variants of both ITS region and RuBisCo spacer were identified in Dongfang No.2 and most of them did not exist in either L. japonica or L. longissima. The unexpected variants may be due to the mutation of ga- metophyte clones occurring during their vegetative amplification.展开更多
Costaria costata(C.Agardh) Saunders is one of common kelps distributed in many coastal areas worldwide;however,in China,no reports have been made on cultivation of the genus.To investigate potential cultivation of the...Costaria costata(C.Agardh) Saunders is one of common kelps distributed in many coastal areas worldwide;however,in China,no reports have been made on cultivation of the genus.To investigate potential cultivation of the species in the northern part of China,trials on isolation and preservation of the gametophytes were conducted using C.costata from Korea;growth and development of the gametophytes were observed.We showed that at 10±1°C,60 μmol m-2s-1 and 12:12 h(L:D),freshly released zoospores settled down within 1 hour,and then developed into the primary cell during the following 2 days.After a vegetative growth phase lasting 6-8 days,female gametophytes became 3-4 times larger in diameter than that of the primary cell,but still remained at a unicellular stage,while male gametophytes divided into 4-10 cells with only a slight change in size.Fertilization occurred within 10 days after the zoospores were released from the sporangia,and the apical and basal tissues of the juvenile sporophyte divided and differentiated into the blade and stipe.Temperature and irradiance influenced gametophytic vegetative growth and developmental patterns.Generally,low irradiance(15 μmol m-2s-1 and 30 μmol m-2s-1) was unfavorable to the induction of fertility,but it enhanced female gametophyte division.The optimal conditions for vegetative growth were 15°C and 30 μmol m-2s-1.After transplantation of the juvenile seedlings and after eight months cultivation,the harvested mature blade reached 194 cm in length and 32.7 cm in width.Our study proves that it is feasible to implement propagation and large scale cultivation of C.costata in northern China.展开更多
Laminaria gametophyte was greatly influenced by light in its growth and development. Using light-emitting diodes (LED) as blue and red light sources, we analyzed the light effect on gametophytes devel- opment of Lamin...Laminaria gametophyte was greatly influenced by light in its growth and development. Using light-emitting diodes (LED) as blue and red light sources, we analyzed the light effect on gametophytes devel- opment of Laminaria japonica Aresch. The gametophytes were obtained from zoospores collected in April, May, July, 2003 and September, 2004. We found that the growth of gametophytes was stimulated by increasing inten- sity of blue light (BL) and red light (RL) illumination, of which BL was obviously stronger than that of RL. The fertilization of gametophytes depended largely on BL, and only sufficient BL illumination could take the repro- ductive effect. In addition, we noticed that there was a significant difference in light responses for gametophytes developed from zoospore collected in different times. For zoospores released in April, under BL1 (73.90 μmol photons/m·s), the unicellular female gametophytes and multi-cellular male gametophytes produced eggs and sperms respectively, and further developed towards sporophytes. However, for gametophytes developed in May, July or September, they became multi-cellular and never formed oogonia or antheridia. It is believed that the Laminaria sporangium maturation stage could affect the gametophytes reaction to BL under laboratory culture conditions. Therefore, cryptochrome- or phototropin-like BL photoreceptors is probably involved in BL-induced development of Laminaria gametophytes.展开更多
基金supported by the National Youth Talent Program(Grant No.A279021801)Key-Area R&D Program of Guangdong Province(Grant No.2022B0202060001)+3 种基金Science and Technology Innovation Team of Shaanxi(Grant No.2021TD-32)the Natural Science Foundation of Shaanxi Province(Grant Nos.2021JM-089,2022JM112 and 2022JQ-162)the Key R&D Project from Yangling Seed Industry Innovation Center(Grant No.K3031322016)Fundamental Research Funds for the Central Universities(Grant No.2452022111)。
文摘As a prerequisite for sexual reproduction,gametophyte development is an interesting process involving cell proliferation,differentiation and specialization.Gametogenesis has been extensively explored in model plants,but the regulatory mechanism of gametophyte development largely remains unknown in Cucurbitaceae species.In present study,we have shown that watermelon ClBBM and ClPLT2,two AP2/ERF transcription factors,participated in both male and female gametophyte development.Clbbm and Clplt2 single mutants resembled wild-type phenotypes in both vegetative and reproductive development.But Clbbm/Clplt2 double mutant showed partial pollen abortion and bore less seeds comparing to WT.Our results indicated that the abnormal pollen grains were caused by premature tapetum degeneration,and reduced seed-set was due to faulty embryo sac development.ClBBM and ClPLT2 were expressed in FG4 embryo sacs,and their transcripts were also detectable in the tapetum and microspore of stage 9 anthers,which was consistent with developmental stages of defective phenotypes observed in double mutant.The expression of genes essential for tapetum development,ClATM1,ClAMS,ClMS1 and ClMS188,was decreased in Clbbm/Clplt2 double mutants.Moreover,the transcriptome analysis indicated that ClBBM and ClPLT2 participated in tapetum and pollen wall development by regulate cell cycle,transmembrane transport,glucan and cellulose metabolic process.Collectively,ClBBM and ClPLT2 were functionally redundant in regulating gametophyte development in watermelon,and their functions differ from their homologous genes in model plant Arabidopsis.
基金This paper is translated from its Chinese version in Scientia Agricultura Sinica.This research was partially supported by the Transcentury Training Program Foundation for the Talents by the Ministry of Education of China to Dr.Chen Jinfeng(30470120)by the National Natural Science Foundation of China(30671419)+2 种基金the National Hi-Tech R&D Program(2004AA241120)the Tang Foundation Cornell-China Scholar Programthe Pickle Seed Research Foundation of Pickle Packers International.The authors sincerely thank Dr.Zhai Huqu,the President of the Chinese Academy of Agricultural Sciences for his support in this research.
文摘An interspecific hybrid F1 of Cucumis hystrix Chakr. x Cucumis sativus L. （NC4406） was used to establish the developmental sequence and to characterize the male and female gametophytes at cytological level for further understanding of the phylogenic relationship and the mechanism of fertility or sterility in the interspecific hybrid F1 The development of male and female gametophytes was studied through meiotic analysis and paraffin section observation technique, respectively. Meanwhile, the fertility level was assessed through hybrid F1 backcrossing to cultivated cucumber 4406. Variable chromosome configurations were observed in the pollen mother cells （PMCs） of hybrid F1 at metaphase Ⅰ , e.g., univalents, bivalents, trivalents, quadravalents, etc. At anaphase Ⅰ and Ⅱ, chromosome lagging and bridges were frequently observed as well, which led to the formation of polyads and only a partial number of microspores could develop into fertile pollen grains （about 23.3%）. Observations of the paraffin sections showed numerous degenerated and abnormal embryo sacs during the development of female gametophytes, and only 40% of the female gametophytes could develop into normal eight-nuclear megaspore. On an average, 22.8 and 6.3 seeds per fruit could be obtained from the reciprocal backcross. The interspecific hybrid F1 of C. hystrix x NC4406 was partially fertile; however, the meiotic behaviors of hybrid F1 showed a high level of intergenomic recombination between C. hystrix and C. sativus chromosomes, which indicated that it plays an important role for introgression of useful traits from C. hystrix into C. sativus.
基金supported in part by the Foundation for Jiangsu Vocational College of Agriculture and Forestry of China(No.2017kj02)the Key Laboratory of Soil Environment and Pollution Remediation,Institute of Soil Science,Chinese Academy of Sciences(No.SEPR2017-10)+2 种基金the National Natural Science Foundation of China(No.21277070)Jiangsu Provincial Innovation Project of China(No.0211001802)the Program for Postgraduates Research and Innovation in Jiangsu Province,China(No.KYLX0052)
文摘Dear Editor, Environmental arsenic (As) contamination of both soil and water systems is a worldwide problem, and As can enter the human food chain through contaminated agricultural products and drinking water and contribute to the development of lung.
文摘Gametophytes of Laminaria japonica were cryopreserved in liquid nitrogen using encapsulation-dehydration with two-step cooling method. Gametophytes cultured at 10℃ and under continuous irradiance of 30 μmol m^-2 s^-1 for 3 weeks were encapsulated in calcium alginate beads. The beads were dehydrated in 0.4 molLl sucrose prepared with seawater for 6 h, desiccated in an incubator set at 10℃ and 70% relative humidity for 4 h, pre-frozen at either -40℃ or -60℃ for 30 min, and stored in liquid nitrogen for 〉24 h. As high as 43% of survival rate was observed when gametophytes were thawed by placing the beads in 40℃ seawater and re-hydrated in 0.05 molL^-1 citrate sodium prepared using 30‰ NaCl 7 d later. More cells of male gametophytes survived the whole procedure in comparison with female gametophytes. The cells of gametophytes surviving the preservation were able to grow asexually and produce morphologically normal sporophytes.
基金Supported by the Shandong Postdoctoral Science Foundation Funded Project(No.2009003027)the National Natural Science Foundation of China(No.31100010)+2 种基金the Key Laboratory of Experimental Marine Biology of Institute of Oceanology,Chinese Academy of Sciences(No.Kf201017)the Qingdao Municipal Science and Technology Project(No.09-1-3-13-jch)the Award Program for Outstanding Young Researchers of Shandong Province(No.BS2009NY018)
文摘A newly identified bacterial disease of kelp(Saccharina japonica) gametophytes was found in clone cultures.It is characterized by swollen gametophyte cells in the early period of infection followed by filamentous fading.An alginolytic marine bacterium referred to as A-1 was isolated from the diseased gametophytes.On the basis of 16S rDNA sequencing and morphological,physiological and biochemical characteristics,the bacterium was identified as a strain of the genus Alteromonas.By testing Koch's postulates,Alteromonas sp.A-1 was further confirmed as the pathogen.The infection process was also investigated using both scanning electron and light microscopy.
文摘This study on the effects of ultrasonic treatment on female gametophytes of Laminariajaponica showed that:1. Ultrasonic treatment had shortening effect on filaments of female gametophytes. Within certainperiod of time, the average length of filamentous female gametophytes was shortened.2. Ultrasonic treatment had emptying effect on cells. The number of empty cells increased with timeof treatment. Ultrasonic treatment had harmful effect on cells.3. Ultrasonic treatment could break down cell walls. The combination of frequency of 20 kHz, out-put of 15 W, 40 s and 60 s of treatment was best for this purpose. After ultrasonic treatment, the regner-ation of female gametophytes into sporophytes was effected. Female gametophytes could not recover aftertoo long period of treatment.
基金Supported by the National Natural Science Foundation of China(Nos.41176135,41206142)the Innovation Program of Chinese Academy of Sciences(No.KSCX2-EW-B-3)the Ministry of Agriculture of China(No.200903030)
文摘The annual life cycle of the brown seaweed Undariapinnatifida （Harvey） Suringer comprises a macroscopic diploid sporophyte stage and a microscopic haploid gametophyte stage. In 2011, an unusual zoospore-derived monoecious gametophyte isolate （designated as line 10-5-3） of U. pinnatifida was observed. To understand this phenomenon, a comprehensive screening of eighty-two previously identified male gametophyte cultures, isolated from three randomly selected cultivars （lines 10, 7, and 5） was performed. Thirty-six of the isolates developed both antheridia and oogonia on the same filamentous fragment in a standard gametogenesis test （SGT： 18℃, 60 umol photons/（m2.s））. Selfing of the monoecious gametophyte or crossing it with a normal male gametophyte both gave rise to morphologically normal sporophytic offspring. However, crossing resulted in a much higher fertilization rate （89.7%）. The hybrid and selfed sporophytic offspring were grown to maturity in flow tanks at an ambient temperature of 10-18℃ over a period of 69 days. Active zoospores were released from both types of mature sporophylls. The majority of these developed into male gametophytes, while 15%-20% developed into the observed monoecious structures on the same filament. Using PCR amplification it was found that all the monoecious gametophyte isolates and the sporophytic offspring resulting from the selfing and crossing lacked the femalelinked microsatellite sequence （a part of the locus Up-AC-2A8, GenBank accession No. AY738602.1）, indicating their male nature. U.pinnatifida is an invasive species in some regions and the implications of the above findings for this species in nature are briefly discussed.
文摘The objective of this study was to examine the eff ects of temperature and light intensity on growth of female gametophytes of Eisenia bicyclis and Ecklonia cava and responses of these female gametophytes to Fe addition and daylength. Female gametophytes of each species were cultured at four temperatures (10, 15, 20, and 25°C) and under a combination of four light intensities (10, 20, 40, and 80 μmol photons/(m^2 ·s)) with two temperatures (15 and 20°C for E i . bicyclis;20 and 25°C for E c . cava ) to clarify their optimal growth conditions. Growth and maturation of female gametophytes of these two species under a combination of five Fe-EDTA concentrations (0, 1, 2, 4, and 8 μmol/L) and three daylengths (10, 12, and 14 h) were also examined. The growth of E i . bicyclis gametophyte was maximal at approximately 15–20°C, 20 μmol photons/(m^2 ·s), Fe-EDTA concentration of 8 μmol/L and daylengths of 12–14 h. While E c . c ava gametophytes showed optimal growth at approximately 20–25°C, 20 μmol photons/(m^2 ·s), Fe- EDTA concentration of 8 μmol/L and daylength of 14 h. Maturation of gametophytes was enhanced at Fe-EDTA concentration of 4 μmol/L for E i . bicyclis and at 2 μmol/L for E c . c ava . In conclusion, optimal growth temperatures and Fe-EDTA concentrations for maturation of E i . bicyclis and E c . c ava gametophytes were diff erent. Higher optimal growth temperature for E c . c ava gametophytes may contribute to its wider geographical distribution compared to E i . bicyclis which has restricted habitats in Korea. This suggests that addition of Fe ion could be used to recover beds of these two species in barren grounds of Korea.
文摘Abstract: Inter-simple sequence repeat (ISSR) analysis was used to assess genetic diversity among 10 pairs of male and female Laminaria gametophytes. A total of 58 amplification loci was obtained from 10 selected ISSR primers, of which 34 revealed polymorphism among the gametophytes. Genetic distances were calculated with the Dice coefficient ranging from 0.006 to 0.223. A dendrogram based on the unweighted pair-group method arithmetic (UPGMA) average showed that most male and female gametophytes of the same species were clustered together and that 10 pairs of gametophytes were divided into four groups. This was generally consistent with the taxonomic categories. The main group consisted of six pairs of gametophytes, which were selected from Laminaria japonica Aresch. by intensive inbreeding through artificial hybridization. One specific marker was cloned, but was not converted successfully into a sequence characterized amplified region (SCAR) marker. Our results demonstrate the feasibility of applying ISSR markers to evaluate Laminaria germplasm diversities.
基金This research was supported by the National Key R&D Program of China(Grant No.2018YFD0901500)the National Natural Science Foundation of China(Grant No.41376136)the Double First-Class Discipline of Fisheries Science of China.
文摘Carbonic anhydrase(CA)was considered to be an important component of carbon concentrating mechanism(CCM)of algae.It was an inducible enzyme.Environmental factors,especially dissolved inorganic carbon and pH,were known to affect CA activity.Effects of inorganic carbon(CO_(2)and HCO_(3)^(-))and pH on CA activity of gametophytes of Saccharina japonica were evaluated in this study.Under high-CO_(2)condition(3%CO_(2)),the activity of external CA(C_(Aext))was significantly decreased(P<0.05)from 33.92 REA/g FW to 27.69 REA/g FW.In contrast,the internal CA(CA_(int))and total CA activities were elevated significantly(p<0.01)from 36.83 REA/g FW to 48.80 REA/g FW,and from 70.75 REA/g FW to 76.49 REA/g FW,respectively.Addition of an appropriate concentration of HCO_(3)^(-)to the medium,CA_(int)and total CA activities were promoted significantly,although the activity of C_(Aext)was inhibited significantly(P<0.05).In higher HCO_(3)^(-)concentration(up to 1680 mg/L),the activity of CA_(int)was promoted to 60.81REA/g FW,which was twice that of the control.Higher activities of CA_(int)and total CA were induced at low pH.At pH 6.0,the highest activities of CA_(int)(110.85 REA/g FW)and total CA(128.17 REA/g FW)were induced.However,at pH 9.0,the activities of CA_(int)and total CA were reduced to the lowest of 23.31REA/g FW and 42.19 REA/g FW,respectively.This is the first report about the successful detection of C_(Aext)activity of gametophytes of S.japonica.The results would provide data for the analysis of Ci acquisition and transport mechanism in S.japonica gametophytes.
基金Supported by the National High Technology Research and Development Program of China(Grant Nos.20060110A4013 and 2006AA100311)National Natural Science Foundation of China(Grant No.40506030)Innovative Key Project of the Chinese Academy of Sciences(Grant No.KZCX2-YW-209)
文摘The life cycle of seaweed Laminaria japonica involves a generation alternation between diploid sporophyte and haploid gametophte.The expression of foreign genes in sporophte has been proved.In this research,the recombinant expression in gametophyte was investigated by particle bombardment with the rt-PA gene encoding the recombinant human tissue-type plasminogen activator(Reteplase),which is a thrombolytic agent for acute myocardial infarction(AMI).Transgenic gametophytes were selected by their resistance to herbicide phosphiothricin(PPT),and proliferated in an established bubble column photo-bioreactor.According to the results from quantitative ELISA,Southern blotting,and fibrin agarose plate assay(FAPA)for bioactivity,it was showed that the rt-PA gene had been inte-grated into the genome of gametophytes of L.japonica,and the expression product showed the ex-pected bioactivity,implying the proper post-transcript modification in haploid gametophyte.
基金中国科学院资助项目，the Project of ScientificInnovation of the Chinese Academy of Sciences，国家科技攻关项目
文摘Effects of various concentrations of two heavy metals, namely Cd and Cu, on gametophytes of Laminariajaponica Aresch were determined by recording morphological changes of gametophytes, determining pH values and the heavy metal content of the culture solution, calculating the germination rate of sporophytes, and observing heavy metal （Cd） distribution using a fluorescence microscope. The results showed that heavy metals damaged the gametophytes, and were even lethal, and that the higher the concentration of heavy metal ions, the greater the injury to gametophytes. Gametophytes could not survive in culture solutions containing more than 100 mg/L Cd and 50 mg/L Cu and were only able to survive in culture solution containing a mixture of Cd and Cu up to a concentration of 10 mg/L, which indicates that gametophytes have a higher tolerance to Cd than Cu and that multiple heavy metal ions in solution markedly aggravate the damage to gametophytes compared with individual heavy metal ions. With increases in the concentration of the heavy metal, the burgeoning rate of sporophytes decreased acutely, and solutions containing multiple heavy metal ions caused even more marked harm to sporophytes than solutions containing a single heavy metal ion, because most sporophytes died in mixed solutions. The pH value of the culture medium dropped immediately at the beginning （the first day） of treatment, increased over the following days, and then decreased again. The pH of culture media containing multiple heavy metal ions showed greater variation than media containing a single heavy metal ion, with the extent of the decrease in pH of culture media containing multiple ions being greatest during the last period of the experiment. With increases in the concentration of heavy metals, the capacity of gametophytes to accumulate these ions increased. The blue fluorescent light emitted by the Cd-and Cd-binding protein complex existing in gametophytes in media containing different concentrations of Cd showed clearly the distribution of the ion in gametophytes and the results obtained were consistent with distribution determined using other methods. All results of the present study showed that gametophytes of L. japonica play a remarkable role as heavy metal decontaminators, especially with regard to Cd.
基金Supported by Project of Jiangsu Provincial Department of Education (JHZD06-7)Qing Lan Project of Colleges and Universities in Jiangsu Province (2008 No.30)~~
文摘[Objective] Pollen mother cell miosis and male gametophyte development of pumpkin were observed in this study, to provide some cytological basis for pumpkin anther or microspore culture. [Method] Ehrlich＇s hematoxylin staining-methyl salicylate clearing technique was used for observation and research of the variation of cell structure and chromosomal behavior during pollen mother cell miosis and male gametophyte development of ‘Tianhong＇ pumpkin. [Result] The meiosis in pollen moth- er cells of pumpkin was simultaneous cytokinesis. In the process of nuclear division, nuclear membrane and nucleolus of pumpkin pollen mother cells gradually disappeared in the metaphase I and reappeared in telophase I , phragmoplast formed between the two generated crescent-shaped nuclei without cell wall, the phragmoplast gradually disappeared in the metaphase II and reappeared in telophase II. Phragmoplast spread outward from the center of spindle during the second division was connected with that formed on the central interface of two nuclei during the first division, cell wall of microspores generated from periphery to center. Most of the tetrads contained four sub-cells while a few contained extra small cells. During the period of uniuclete microspore at periphery, the single nucleolus split into 2-3 or more small nucleoli, mature pollen grain was two-celled. Mononucleate pollen cells were mostly appeared in the flower buds with length of 1.0-2.0 cm, which could be used as an important indicator to collect materials for anther or microspore culture. [Conclusion] This study laid the foundation for research of the cytogenetics of pumpkin.
基金Supported by the National Key R&D Program of China(No.2018YFD0901500)the Shanghai Sailing Program(No.17YF1407900),the National Natural Science Foundation of China(No.41576163)the Ocean Public Welfare Scientifi c Research Project,China(No.201205010)。
文摘Abstract Ulva linza is one of the species that causes green tides in the Yellow Sea,China.Due to the diffi culties in chromosomal preparation,the large numbers of chromosomes,and their relatively small sizes,there have been no reported studies on Ulva macroalgae chromosomes.The karyotypes and chromosomes in U.linza were observed after a series of treatments.The chromosomes were pretreated with 0.1%colchicine for 12 h and then mixed with enzymes.The samples were dropped from 30 cm height onto glass slides,which spread out the surface coat.These pretreatments were the optimal chromosomes preparation treatments.The prepared chromosomes were stained with 4',6-diamidino-2-phenylindole(DAPI),which is a fl uorescent probe sensitive and specifi c to DNA.The chromosome number in the haploid male and female gametophytes was n=9,and was 2 n=18 in the diploid sporophytes.The female gametophyte chromosomes were between 0.804 and 2.292μm in size,the male gametophyte chromosomes were between 0.917 and 2.916μm,and the sporophyte chromosomes were between 0.912 and 2.167μm.The relative sizes of the chromosomes were used to analyze the karyotypes of the female and male gametophyte chromosomes.The results provide a solid foundation for the basic technique that can be used to localize molecular markers of Ulva chromosomes.
文摘Eighteen gametophytes includingL. japonica, L. ochotensisandL. longissima, were verified with random amplified polymorphic DNA (RAPD) technique. Eighteen ten-base primers were chosen from 100 primers selected for final amplification test. Among the total of 205 bands amplified, 181 (88.3%) were polymorphic. The genetic distance among different strains ranged from 0.072 to 0.391. The dendrogram constructed by unweighted pair-group method with arithmetic (UPGMA) method showed that the female and male gametophytes of the same cell lines could be grouped in pairs respectively. It indicated that RAPD analysis could be used not only to distinguish different strains ofLaminaria, but also to distinguish male and female gametophyte within the same cell lines. There is ambiguous systematic relationship if judged merely by the present data. It seems that the use of RAPD marker is limited to elucidation of the phylogenetic relationship among the species ofLaminaria.
文摘Unicellular gametophytes of Undaria pinnatifida (Harv.) Sur. were isolated in Qingdao, P. R. China in April 1993 and in Tokushima, southem Japan in March 1995. Diferent intraspecific crossings by using unicellular male and female gametophytes were successfully undertaken in Sept. of 1995 in Qingdao.Sporophytes were transplanted to two different locations for open an cultivation. One was at Zhanqiao (ZQ) Bay where the water current was slower than that of another location - Taipingjiao(TPJ). A total of218 adult sporophytes were harvested on January 12 and 18 from TPJ and ZQ repectively. For eacncombination, 10 sporophytes un cultivated. Analysis of the mprpholgical characteristics of adult spprophytes indicated that the longest length between two bases of the serration of pinnate bldes (W2) is a morphological characteristic that can be transferred from the parent plant to the next generations regardless of environmental variations. There was evidence that W2 was apparently determined by sex-linked factors, i.e, by male parental grametophyte.. Sporophytes from certain crossing combinations showed more vigorous growth than those from other crossing combinations. It is therrfore possible to select grametophyte strains which can be used as parental gametophytes for the seeding production of sporophytes with more vigorous growth within shorter cultivation period. The morphology of hybrids from a Qingdao strain and a Tokushima cultivated strain resembled that of both parental plants in frond feaures (wrinkled or smooth) and W2. Sporophyll formation also varied with strains. The fact that adult sporophytes resulting from the same crossing combinations have identical morphological characteristics under the same environmental conditions indicates the possibility of a new way to select strains which are expected to be ideal for commerical production by purposely selecting, propagating, and seeding unicellular gametophytes for sporeing production through freeliving techniques of gametophytes.
基金Under the auspices of National Key Research and Development Program of China(No.2016YFC0500300)National Natural Science Foundation of China(No.41675153)
文摘Suitable habitat is vital for the survival and restoration of a species.Understanding the suitable habitat range for lycophytes and ferns is prerequisite for effective species resource conservation and recovery efforts.In this study, we took Athyrium brevifrons as an example, predicted its suitable habitat using a Maxent model with 67 occurrence data and nine environmental variables in Northeast China.The area under the curve(AUC) value of independent test data, as well as the comparison with specimen county areal distribution of A.brevifrons exhibited excellent predictive performance.The type of environmental variables showed that precipitation contributed the most to the distribution prediction, followed by temperature and topography.Percentage contribution and permutation importance both indicated that precipitation of driest quarter(Bio17) was the key factor in determining the natural distribution of A.brevifrons, the reason could be proved by the fern gametophyte biology.The analysis of high habitat suitability areas also showed the habitat preference of A.brevifrons: comparatively more precipitation and less fluctuation in the driest quarter.Changbai Mountains, covering almost all the high and medium habitat suitability areas, provide the best ecological conditions for the survival of A.brevifrons, and should be considered as priority areas for protection and restoration of the wild resource.The potential habitat suitability distribution map could provide a reference for the sustainable development and utilisation of A.brevifrons resource, and Maxent modelling could be valuable for conservation management planning for lycophytes and ferns in Northeast China.
文摘The cultivation of the first filial generation of monoploid gametophyte clones of different Laminaria species (hybrid Laminaria) is an effective way of utilizing heterozygous vigor (heterosis). A female gametophyte clone of L. japonica and a male gametophyte clone of L. longissima were hybridized, generating Dongfang No.2 hybrid Laminaria. The parentage of this hybrid Laminaria was determined using AFLP of total DNA, SNP of the ITS region of ribosomal RNA transcription unit and microsatellite DNA variation at two loci. In addition to 167 AFLP bands shared by Dongfang No.2, L. japonica and L. longissima, Dongfang No.2 hybrid Laminaria shared another 70 and 55 bands with L. japonica and L. longissima, respectively, which were obviously more than 11 bands shared by L. japonica and L. longissima. Dongfang No.2 held both ‘T’ and ‘C’ at position 847 of the ITS region, while ‘T’ at this position was specific for L. japonica and ‘C’ for L. longissima, respectively. Dongfang No.2 also held the microsatellite DNA alleles of two parents together at two microsatellite DNA marker loci. These observations clearly proved that Dongfang No.2 is a true hybrid of L. japonica and L. longissiuma. Unfortunately, the origin of the chloroplast of Dongfang No.2 was not determined based on the variation of RuBisCo spacer. More sequence variants of both ITS region and RuBisCo spacer were identified in Dongfang No.2 and most of them did not exist in either L. japonica or L. longissima. The unexpected variants may be due to the mutation of ga- metophyte clones occurring during their vegetative amplification.
基金Supported by the Shandong Agriculture Seedstocks Projectthe Knowledge Innovation Program of the Chinese Academy of Sciences(No.KSCX2-YW-N-247-02)
文摘Costaria costata(C.Agardh) Saunders is one of common kelps distributed in many coastal areas worldwide;however,in China,no reports have been made on cultivation of the genus.To investigate potential cultivation of the species in the northern part of China,trials on isolation and preservation of the gametophytes were conducted using C.costata from Korea;growth and development of the gametophytes were observed.We showed that at 10±1°C,60 μmol m-2s-1 and 12:12 h(L:D),freshly released zoospores settled down within 1 hour,and then developed into the primary cell during the following 2 days.After a vegetative growth phase lasting 6-8 days,female gametophytes became 3-4 times larger in diameter than that of the primary cell,but still remained at a unicellular stage,while male gametophytes divided into 4-10 cells with only a slight change in size.Fertilization occurred within 10 days after the zoospores were released from the sporangia,and the apical and basal tissues of the juvenile sporophyte divided and differentiated into the blade and stipe.Temperature and irradiance influenced gametophytic vegetative growth and developmental patterns.Generally,low irradiance(15 μmol m-2s-1 and 30 μmol m-2s-1) was unfavorable to the induction of fertility,but it enhanced female gametophyte division.The optimal conditions for vegetative growth were 15°C and 30 μmol m-2s-1.After transplantation of the juvenile seedlings and after eight months cultivation,the harvested mature blade reached 194 cm in length and 32.7 cm in width.Our study proves that it is feasible to implement propagation and large scale cultivation of C.costata in northern China.
基金This research was supported by the NSFC (No.40376049) and Japan Society for the Promotion of Science (The Invitation Fellowship for Research in Japan 2002)
文摘Laminaria gametophyte was greatly influenced by light in its growth and development. Using light-emitting diodes (LED) as blue and red light sources, we analyzed the light effect on gametophytes devel- opment of Laminaria japonica Aresch. The gametophytes were obtained from zoospores collected in April, May, July, 2003 and September, 2004. We found that the growth of gametophytes was stimulated by increasing inten- sity of blue light (BL) and red light (RL) illumination, of which BL was obviously stronger than that of RL. The fertilization of gametophytes depended largely on BL, and only sufficient BL illumination could take the repro- ductive effect. In addition, we noticed that there was a significant difference in light responses for gametophytes developed from zoospore collected in different times. For zoospores released in April, under BL1 (73.90 μmol photons/m·s), the unicellular female gametophytes and multi-cellular male gametophytes produced eggs and sperms respectively, and further developed towards sporophytes. However, for gametophytes developed in May, July or September, they became multi-cellular and never formed oogonia or antheridia. It is believed that the Laminaria sporangium maturation stage could affect the gametophytes reaction to BL under laboratory culture conditions. Therefore, cryptochrome- or phototropin-like BL photoreceptors is probably involved in BL-induced development of Laminaria gametophytes.
