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
[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
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
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
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
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
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
In China, the total annual production of cultured Laminaria japonica reached half million tons in dry weight there few years. The routine sporeling culture technique conducted in the greenhouse took at least three and...In China, the total annual production of cultured Laminaria japonica reached half million tons in dry weight there few years. The routine sporeling culture technique conducted in the greenhouse took at least three and half months. In such a case, sometimes the sporelings died within a few days due to destructive diseases. In Order to overcome the mentioned problems, a new sporeling culture technique, the clone technique, is developed. The method includes three stead: (1) Gametophyte clone culture. The spores and the gametophytes are cloned in flasks under favorable environments. (2) Sporeling cul ture. Male and female clones are crushed and spread onto a frame to allow the gametophytes to attach to the substrata. The frames are cultured in tanks, and the sporophytes reach 1 cm in length within one and a half months. (3) Outgrowing of the plant. The frames are put in the open sea when seawater temperature decreased to 20℃. After one month, the sporelings are large enough to be transplanted. It is concluded that the clone technique has the following advantages: (1)Large amount of clones can be produced in a short period of time. (2) Clone seeding method makes it free from the biological rhythm, one can seed the plant anytime all the year round. (3) It takes only one and a half months to complete the process of sporeling cultivation in the greenhouse. At present, this technique is used in the breeding of new strains of Laminaria.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
The red alga Gracilariopsis lemaneiformis(Bory) is an economically valuable macroalgae. As a means to identify the sex of immature Gracilariopsis lemaneiformis, the amplifi ed fragment length polymorphism(AFLP) techni...The red alga Gracilariopsis lemaneiformis(Bory) is an economically valuable macroalgae. As a means to identify the sex of immature Gracilariopsis lemaneiformis, the amplifi ed fragment length polymorphism(AFLP) technique was used to search for possible sex- or phase-related markers in male gametophytes, female gametophytes, and tetrasporophytes, respectively. Seven AFLP selective amplifi cation primers were used in this study. The primer combination E-TG/M-CCA detected a specifi c band linked to male gametophytes. The DNA fragment was recovered and a 402-bp fragment was sequenced. However, no DNA sequence match was found in public databases. Sequence characterized amplifi ed region(SCAR) primers were designed from the sequence to test the repeatability of the relationship to the sex, using 69 male gametophytes, 139 female gametophytes, and 47 tetrasporophytes. The test results demonstrate a good linkage and repeatability of the SCAR marker to sex. The SCAR primers developed in this study could reduce the time required for sex identifi cation of Gracilariopsis lemaneiformis by four to six months. This can reduce both the time investment and number of specimens required in breeding experiments.展开更多
Florogenesis is one of the most complicated and interesting processes in the nature. This process involves developmental, physiological and molecular events leading to transformation from vegetative to reproductive ph...Florogenesis is one of the most complicated and interesting processes in the nature. This process involves developmental, physiological and molecular events leading to transformation from vegetative to reproductive phase for optimal seed production and the continuation of species. The basic knowledge about flowering processes, male and female sexual systems support basic and applied research and breeding programs. Most of the onion varieties from India are short day varieties, more diverse than other exotic germplasm and useful as a source of new alleles for supporting breeding programs. The present investigation was focused to study for the first time florogenesis process by scanning electron study for the first time florogenesis process by scanning electron microscopy (SEM) and development of female gametophytes by light microscopy in order to acquire basic knowledge useful for optimizing in vitro process to produce gynogenic haploid to support and speed breeding program in short-day onion Allium cepa L. cv. Krishnapuram (KP) or Bangalore Rose. This study revealed that shoot primordium differentiated into inflorescence meristem in the month of December, while seeds were planted in the field in the September. The individual florets are preceded by a variying number of floral initials. The female gametophyte developed from chalazal side megaspore. The embryo sac development is a bisporic Allium type showing short-lived antipodals. The histological study suggests that the use of big or preanthesis flower buds with embryo sac for production of gynogenic haploids to support breeding program in onion cv. Krishnapuram (KP). However further studies are needed for confirmation of this observation.展开更多
The microsporogenesis and male gametophyte development of Euphorbia pulcherrima were studied with paraffin section method. The results showed that the anther of Euphorbia pulcherrima had four chambers. The development...The microsporogenesis and male gametophyte development of Euphorbia pulcherrima were studied with paraffin section method. The results showed that the anther of Euphorbia pulcherrima had four chambers. The development of anther wall was dicotyledonous type; its wall consisted of epidermis, endothecium, middle layer and tapetum. The mature male gametophyte development experienced seven major stages as the followings： archesporial cells, aporgenous cells, microspore mother cells, tetrads of microspores （tetrahedral）, uninucleate microspores, 2-uncleate pollens and 2-celled pollen stage. The anther wall varied with the pollen development： the epidermis of anther ruptures at the anther mature stage; the middle layer disappears at tetrad stage; the endothelium became fibrous layer at the anther mature stage; the tapetum （glandular） was the most development at the microspore mother cells stage and disappeared at anther mature stage. The abnormal anther development was observed simultaneously.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
Pyropia haitanensis(T.J.Chang et B.F.Zheng) N.Kikuchi et M.Miyata( Porphyra haitanensis) is an economically important genus that is cultured widely in China.P.haitanensis is cultured on a larger scale than Pyropia yez...Pyropia haitanensis(T.J.Chang et B.F.Zheng) N.Kikuchi et M.Miyata( Porphyra haitanensis) is an economically important genus that is cultured widely in China.P.haitanensis is cultured on a larger scale than Pyropia yezoensis,making up an important part of the total production of cultivated Pyropia in China.However,the majority of molecular mechanisms underlying the physiological processes of P.haitanensis remain unknown.P.haitanensis could utilize inorganic carbon and the sporophytes of P.haitanensis might possess a PCK-type C 4-like carbon-fixation pathway.To identify micro RNAs and their probable roles in sporophyte and gametophyte development,we constructed and sequenced small RNA libraries from sporophytes and gametophytes of P.haitanensis.Five micro RNAs were identified that shared no sequence homology with known micro RNAs.Our results indicated that P.haitanensis might posses a complex s RNA processing system in which the novel micro RNAs act as important regulators of the development of different generations of P.haitanensis.展开更多
The aim of this study is to follow each development stage of inflorescence in order to understand the biological feature of flowering and the development of male gametophyte in Anthurium andreanum “Arizona' and ...The aim of this study is to follow each development stage of inflorescence in order to understand the biological feature of flowering and the development of male gametophyte in Anthurium andreanum “Arizona' and to try to find the optimum conditions for its pollination. The methods of dissection and paraffin section were adopted to examine the structural characteristics of anthurium’s tiny floret and the development of the microspore. All the florets of the anthurium arrange on the rhachis helically sub- tended by a colorful bract. Each tiny floret has one gynoecium, four tepals and four stamina. The bract and the florets show different colors during the whole blooming period. The ovary is bicarpellary and has two locules, each of which has one anatropous ovule. The placenta is of a central placentation type. The stylar canal cells not only can produce the secretory mucilage but also can release their own cytoplasm caused by their self-disintegration before the pistil reaches its maturity. The wall of the anther is composed of four layers: epidermis, endothecium, middle layer and tapetum. The tapetal cells and the middle layers’ cells degenerated completely dur- ing meiosis of microsporocytes. The pollen grains were 2-celled at the time of anther dehiscence. Early morning, when the inflores- cences stay at their fifth development stage, is the optimum opportunity for pistil to get pollen grains. The pollen-collection should be done at the end of the seventh stage.展开更多
There were four microsporanges in the mature anther of B. rossica Fedtsch. et Flerov. The development of the anther wall belonged to dicotyledonous type. The tapetum cell was dichotypic. Simultaneous cytocinesis in th...There were four microsporanges in the mature anther of B. rossica Fedtsch. et Flerov. The development of the anther wall belonged to dicotyledonous type. The tapetum cell was dichotypic. Simultaneous cytocinesis in the microspore mother cells formed tetrad by reduction division and the array of microspore tetrads was different. There were tetrahedron type and bilateral symmetry type. The male gametophyte was 2-cell. The mature pollen was global and tricolporate. All those results were investigated under both light microscope （LM） and electron microscope （EM）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 rec...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
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.
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.
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.
基金Key Project, the Ninth Five-Year Plan of China! 96-008-01-02-06Project of Bio-Engineering Center of China! SSTC 96-C01-05-01
文摘In China, the total annual production of cultured Laminaria japonica reached half million tons in dry weight there few years. The routine sporeling culture technique conducted in the greenhouse took at least three and half months. In such a case, sometimes the sporelings died within a few days due to destructive diseases. In Order to overcome the mentioned problems, a new sporeling culture technique, the clone technique, is developed. The method includes three stead: (1) Gametophyte clone culture. The spores and the gametophytes are cloned in flasks under favorable environments. (2) Sporeling cul ture. Male and female clones are crushed and spread onto a frame to allow the gametophytes to attach to the substrata. The frames are cultured in tanks, and the sporophytes reach 1 cm in length within one and a half months. (3) Outgrowing of the plant. The frames are put in the open sea when seawater temperature decreased to 20℃. After one month, the sporelings are large enough to be transplanted. It is concluded that the clone technique has the following advantages: (1)Large amount of clones can be produced in a short period of time. (2) Clone seeding method makes it free from the biological rhythm, one can seed the plant anytime all the year round. (3) It takes only one and a half months to complete the process of sporeling cultivation in the greenhouse. At present, this technique is used in the breeding of new strains of Laminaria.
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.
基金Supported by the"Twelfth Five-Year-Plan"of National Science and Technology for the Rural Development in China(No.2012AA10A411)the Public Welfare Project of the Ministry of Agriculture of China(No.200903030)
文摘The red alga Gracilariopsis lemaneiformis(Bory) is an economically valuable macroalgae. As a means to identify the sex of immature Gracilariopsis lemaneiformis, the amplifi ed fragment length polymorphism(AFLP) technique was used to search for possible sex- or phase-related markers in male gametophytes, female gametophytes, and tetrasporophytes, respectively. Seven AFLP selective amplifi cation primers were used in this study. The primer combination E-TG/M-CCA detected a specifi c band linked to male gametophytes. The DNA fragment was recovered and a 402-bp fragment was sequenced. However, no DNA sequence match was found in public databases. Sequence characterized amplifi ed region(SCAR) primers were designed from the sequence to test the repeatability of the relationship to the sex, using 69 male gametophytes, 139 female gametophytes, and 47 tetrasporophytes. The test results demonstrate a good linkage and repeatability of the SCAR marker to sex. The SCAR primers developed in this study could reduce the time required for sex identifi cation of Gracilariopsis lemaneiformis by four to six months. This can reduce both the time investment and number of specimens required in breeding experiments.
文摘Florogenesis is one of the most complicated and interesting processes in the nature. This process involves developmental, physiological and molecular events leading to transformation from vegetative to reproductive phase for optimal seed production and the continuation of species. The basic knowledge about flowering processes, male and female sexual systems support basic and applied research and breeding programs. Most of the onion varieties from India are short day varieties, more diverse than other exotic germplasm and useful as a source of new alleles for supporting breeding programs. The present investigation was focused to study for the first time florogenesis process by scanning electron study for the first time florogenesis process by scanning electron microscopy (SEM) and development of female gametophytes by light microscopy in order to acquire basic knowledge useful for optimizing in vitro process to produce gynogenic haploid to support and speed breeding program in short-day onion Allium cepa L. cv. Krishnapuram (KP) or Bangalore Rose. This study revealed that shoot primordium differentiated into inflorescence meristem in the month of December, while seeds were planted in the field in the September. The individual florets are preceded by a variying number of floral initials. The female gametophyte developed from chalazal side megaspore. The embryo sac development is a bisporic Allium type showing short-lived antipodals. The histological study suggests that the use of big or preanthesis flower buds with embryo sac for production of gynogenic haploids to support breeding program in onion cv. Krishnapuram (KP). However further studies are needed for confirmation of this observation.
文摘The microsporogenesis and male gametophyte development of Euphorbia pulcherrima were studied with paraffin section method. The results showed that the anther of Euphorbia pulcherrima had four chambers. The development of anther wall was dicotyledonous type; its wall consisted of epidermis, endothecium, middle layer and tapetum. The mature male gametophyte development experienced seven major stages as the followings： archesporial cells, aporgenous cells, microspore mother cells, tetrads of microspores （tetrahedral）, uninucleate microspores, 2-uncleate pollens and 2-celled pollen stage. The anther wall varied with the pollen development： the epidermis of anther ruptures at the anther mature stage; the middle layer disappears at tetrad stage; the endothelium became fibrous layer at the anther mature stage; the tapetum （glandular） was the most development at the microspore mother cells stage and disappeared at anther mature stage. The abnormal anther development was observed simultaneously.
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.
基金Supported by the National High Technology Research and Development Program of China(863 Program)(No.2012AA10A406)the National Natural Science Foundation of China(Nos.41176134,41306151)the China Strategic Leading Special Science and Technology Academy(No.XDA11020404)
文摘Pyropia haitanensis(T.J.Chang et B.F.Zheng) N.Kikuchi et M.Miyata( Porphyra haitanensis) is an economically important genus that is cultured widely in China.P.haitanensis is cultured on a larger scale than Pyropia yezoensis,making up an important part of the total production of cultivated Pyropia in China.However,the majority of molecular mechanisms underlying the physiological processes of P.haitanensis remain unknown.P.haitanensis could utilize inorganic carbon and the sporophytes of P.haitanensis might possess a PCK-type C 4-like carbon-fixation pathway.To identify micro RNAs and their probable roles in sporophyte and gametophyte development,we constructed and sequenced small RNA libraries from sporophytes and gametophytes of P.haitanensis.Five micro RNAs were identified that shared no sequence homology with known micro RNAs.Our results indicated that P.haitanensis might posses a complex s RNA processing system in which the novel micro RNAs act as important regulators of the development of different generations of P.haitanensis.
基金Supported by the Graduate Students’ Research Foundation of Beijing Forestry University
文摘The aim of this study is to follow each development stage of inflorescence in order to understand the biological feature of flowering and the development of male gametophyte in Anthurium andreanum “Arizona' and to try to find the optimum conditions for its pollination. The methods of dissection and paraffin section were adopted to examine the structural characteristics of anthurium’s tiny floret and the development of the microspore. All the florets of the anthurium arrange on the rhachis helically sub- tended by a colorful bract. Each tiny floret has one gynoecium, four tepals and four stamina. The bract and the florets show different colors during the whole blooming period. The ovary is bicarpellary and has two locules, each of which has one anatropous ovule. The placenta is of a central placentation type. The stylar canal cells not only can produce the secretory mucilage but also can release their own cytoplasm caused by their self-disintegration before the pistil reaches its maturity. The wall of the anther is composed of four layers: epidermis, endothecium, middle layer and tapetum. The tapetal cells and the middle layers’ cells degenerated completely dur- ing meiosis of microsporocytes. The pollen grains were 2-celled at the time of anther dehiscence. Early morning, when the inflores- cences stay at their fifth development stage, is the optimum opportunity for pistil to get pollen grains. The pollen-collection should be done at the end of the seventh stage.
文摘There were four microsporanges in the mature anther of B. rossica Fedtsch. et Flerov. The development of the anther wall belonged to dicotyledonous type. The tapetum cell was dichotypic. Simultaneous cytocinesis in the microspore mother cells formed tetrad by reduction division and the array of microspore tetrads was different. There were tetrahedron type and bilateral symmetry type. The male gametophyte was 2-cell. The mature pollen was global and tricolporate. All those results were investigated under both light microscope （LM） and electron microscope （EM）
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.
