The doubled haploid(DH)technique accelerates homozygosity by inducing chromosome doubling in haploid embryos derived from hybrid plants.This approach offers significant advantages over conventional rice breeding metho...The doubled haploid(DH)technique accelerates homozygosity by inducing chromosome doubling in haploid embryos derived from hybrid plants.This approach offers significant advantages over conventional rice breeding methods by shortening the breeding cycle and enabling rapid development of pure homozygous lines.Anther culture(AC)has been established as an efficient and successful method for producing DH plants via androgenesis in rice.However,despite its success in japonica rice.展开更多
Carotenoids are lipophilic isoprenoid pigments with essential roles in plants.While the cultivated allotetraploid cottons exhibit distinct mature anther coloration—yellow in Gossypium barbadense versus predominantly ...Carotenoids are lipophilic isoprenoid pigments with essential roles in plants.While the cultivated allotetraploid cottons exhibit distinct mature anther coloration—yellow in Gossypium barbadense versus predominantly white in G.hirsutum—the genetic basis of this divergence remains unclear.The purpose of this study was to identify the genetic basis of anther-color variation in cotton(Gossypium)species.We firstly identified carotenoids as the primary pigments underlying yellow-anthers coloration.Comparative transcriptomics of anthers revealed that the carotenoid biosynthesis gene GbPSY4 was expressed as a key regulator in G.barbadense.Functional validation via tissue-specific expression,subcellular localization,in vivo enzymatic assays,and virus-induced gene silencing confirmed its role in carotenoid biosynthesis and yellow pigmentation.Genome-wide association studies in a G.hirsutum population revealed GhPSY4_At,an ortholog of GbPSY4,as the causal gene of anther-color variation.We conclude that PSY4-regulated carotenoid biosynthesis governs yellow pigmentation.Furthermore,a finding that G.hirsutum accessions with yellow anthers showed greater pollen viability under high-temperature stress than those with white anthers suggests that the same pathway that governs yellow pigmentation influences heat tolerance.PSY4 is a promising target for breeding stress-tolerant cotton varieties.展开更多
Recent studies have shown that lipid metabolism is a key factor affecting anther development and male fertility.However,how plants regulating the metabolic balance of multiple lipids to ensure proper anther developmen...Recent studies have shown that lipid metabolism is a key factor affecting anther development and male fertility.However,how plants regulating the metabolic balance of multiple lipids to ensure proper anther development and male fertility remains unclear.Analyzing lipid molecules related to anther fertility and genes responsible for their biosynthesis is crucial for understanding the physiological significance of lipid metabolism in crop fertility.In this study,we compared the transcriptome and the composition and content of lipids in anthers of two upland cotton(Gossypium hirsutum) materials,Shida 98(WT) and its nearly-isogenic male sterile line Shida 98A(MS).Transcriptomics analysis identified many differentially expressed genes(DEGs) between the two materials,with the genes of the alpha-linolenic acid metabolism pathway being the most significantly associated with the male sterility phenotype.Investigations on lipids revealed that the MS anthers over-accumulated free fatty acids(FFAs),phosphatidic acid(PA),mono-and di-galactosyldiacylglycerol(MGDG and DGDG),and had a decreased content of triacylglycerol(TAG),which was closely related to the abnormal metabolism of alpha-linolenic acid(C18:3);therefore,the major lipids containing C18:3-acyl chains,such as PA,MGDG,DGDG,and TAG,are proposed to play a major role in cotton anther development.We also showed that an excessive level of MGDG and DGDG caused jasmonic acid(JA) overaccumulation in MS anthers,which in turn inhibited the expression of GhFAD3 and consequently reduced the C18:3 content,presumably via a feedback regulation mechanism,ultimately affecting plant fertility.Together,our results revealed the importance of a balanced lipid metabolism in regulating the development of cotton anther and pollen and consequently male fertility.展开更多
Carotenoids are natural pigments that are widely distributed in the flowers,fruits,and seeds of many plant species.These compounds not only endow diverse colors but also exhibit antioxidant,immune-modulatory,anti-agin...Carotenoids are natural pigments that are widely distributed in the flowers,fruits,and seeds of many plant species.These compounds not only endow diverse colors but also exhibit antioxidant,immune-modulatory,anti-aging,and photoprotective properties.Although carotenoid metabolism has been studied extensively in microbial and plant science,the genetic mechanisms underlying carotenoid metabolism in cotton remain underexplored.Here,we isolated gene GbDYA that regulates a dark-yellow anther color by map-based cloning using a BC1F1 population derived from a cross of Gossypium barbadense acc.Hai7124 with dark-yellow anthers and G.hirsutum acc.TM-1 with light-yellow anthers backcrossed with TM-1.GbDYA encodes phytoene synthase,a key rate-limiting enzyme in the carotenoid biosynthesis pathway.A long terminal repeat retrotransposon in the first exon of GhDYA(an ortholog of GbDYA in G.hirsutum acc.TM-1)caused loss of function and led to the light-yellow anther color.GbDYA is predominantly expressed in the early stages of anther development.Transcriptome,RT-qPCR and KEGG enrichment analyses revealed that GbDYA influences the synthesis and accumulation of carotenoids in anthers by modulating expression of key genes in the carotenoid biosynthesis pathway.Integrated transcriptomic and metabolomic analyses indicated that the accumulation of lutein,violaxanthin,antherxanthin,cryptoxanthin,zeaxanthin,andβ-carotene contributed to yellow coloration of anthers.Dual-luciferase and yeast one-hybrid assays confirmed that transcription factor GbMYB105(GB_A11G3511)binds to the promoter of GbDYA and activates its expression.High-temperature stress treatment indicated that carotenoids accumulation in anthers enhances pollen antioxidant activity.This study unravels the role of GbDYA in conferring the anther coloration,and provides the potential utilization by modulating accumulation of carotenoids in anthers to enhance pollen viability in high-temperature tolerance breeding in cotton.展开更多
Anther is a key male reproductive organ that is essential for the plant life cycle,from the sporophyte to the gametophyte generation.To explore the isoform-level transcriptional landscape of developing anthers in maiz...Anther is a key male reproductive organ that is essential for the plant life cycle,from the sporophyte to the gametophyte generation.To explore the isoform-level transcriptional landscape of developing anthers in maize(Zea mays L.),we analyzed Iso-Seq data from anthers collected at 10 developmental stages,together with strand-specific RNA-seq,CAGE-seq,and PAS-seq data.Of the 152,026 high-confidence full-length isoforms identified,68.8%have not been described;these include 22,365 isoforms that originate from previously unannotated loci and 82,167 novel isoforms that originate from annotated protein-coding genes.Using our newly developed strategy to detect dynamic expression patterns of isoforms,we identify 13,899 differentially variable regions(DVRs);surprisingly,1275 genes contain more than two DVRs,revealing highly efficient utilization of limited genic regions.We identify 7876 long non-coding RNAs(lncRNAs)from 4098 loci,most of which were preferentially expressed during cell differentiation and meiosis.We also detected 371 long-range interactions involving intergenic lncRNAs(lincRNAs);interestingly,243 were lincRNA-gene ones,and the interacting genes were highly expressed in anthers,suggesting that many potential lncRNA regulators of key genes are required for anther development.This study provides valuable resources and fundamental information for studying the essential transcripts of key genes during anther development.展开更多
Water scarcity is an escalating global challenge that severely threatens productivity and reproductive success in crops,particularly in drought-sensitive species such as Capsicum annuum L.Although deficit irrigation s...Water scarcity is an escalating global challenge that severely threatens productivity and reproductive success in crops,particularly in drought-sensitive species such as Capsicum annuum L.Although deficit irrigation strategies are widely recommended to enhance water use efficiency,knowledge remains limited regarding their interactions with soil amendments such as biochar and the consequent impacts on reproductive traits.This study aimed to evaluate the combined effects of deficit irrigation strategies and biochar application on pollen viability and morphology in Capsicum annuum.The experiment was conducted under full,partial,and deficit irrigation regimes with and without biochar treatment,following a randomized block design.The primary parameters examined were pollen viability(viable,semi-viable,and non-viable rates),anther width and length,and pollen width and length.Microscopic measurements and statistical analyses(p≤0.05)revealed significant effects of both irrigation regimes and biochar applications.Under deficit irrigation,viable,semi-viable,and non-viable pollen rates were 29.84%,32.95%,and 37.21%,respectively,whereas the highest viable pollen rate was observed under full irrigation.In partial irrigation,viable pollen accounted for 31.67%,semi-viable for 38.81%,and non-viable for 29.49%.In plots treated with biochar under partial irrigation,anther width(1700.89μm),anther length(3805.34μm),pollen width(26.93μm),and pollen length(37.42μm)reached the highest values,while the lowest values were recorded in deficit irrigation plots without biochar.These findings emphasize the importance of integrating biochar into irrigation management to mitigate the adverse effects of water stress on pollen development.Nevertheless,further research is needed to clarify the long-term implications of these practices for reproductive success and agricultural sustainability.展开更多
Some influential factors of anther culture were studied preliminarily by conducting anther culture of the restorers of new cytoplasmic male sterile （NER）. Several results were obtain from this experiment and they we...Some influential factors of anther culture were studied preliminarily by conducting anther culture of the restorers of new cytoplasmic male sterile （NER）. Several results were obtain from this experiment and they were listed as follow：① MS cultrure medium with such hormones as 2,4-D 2 mg/L,6-BA 0.5 mg/L, NAA 0.5 mg/L was the best suitable for callus induction of NER. ②The difference of induction rate was significantly different between different plant age groups. From the 110th day to 141th day,the induction rate was increased with the increase of age and the difference of induction rate reached 0.01 significant difference level. The induction rate reached the highest value in the 141th day then it declined gradually. ③The combined use of 2, 4-D and 6-BA with proper increase of 2,4-D was good for inducing callus. ④The green plantlet induction rate of NER was increased when the concentration of 6-BA increased from 2 mg/L to 4 mg/L. Adding ZT from 0.5 mg/L to 2 mg/L. 6-BA would led 2.47% increase of green plantlet olantlet induction rate.展开更多
In this study, through vitro culturing anthers of 7 F1 progenies of early Japonica rice in cold region on medium with different Fe2+ contents, it was found that Fe2+ content generated greater impacts on the induction ...In this study, through vitro culturing anthers of 7 F1 progenies of early Japonica rice in cold region on medium with different Fe2+ contents, it was found that Fe2+ content generated greater impacts on the induction rate and green plantlet differentiation. The result demonstrated that if Fe2+ increased from 32 to 40 mg/kg, the induction rate of early Japonica rice anther culture in N6 culture media was more then 1.4 times higher than that in N6 culture media containing 5.6 mg/kg Fe2+. In this concentration range, the induction rate increased with the increase of Fe2+ content, while if the concentration was over this concentration range, the induction rate decreased with the increase of Fe2+, showing single peak distribution. When the Fe2+ was 40 mg/kg in differentiation medium, the differentiation rate decreased dramatically. The green plantlet differentiations of callus which were induced on culture media containing 32-40 mg/kg Fe2+ were different, when they were cultured on MS culture media, and 85.7% materials could increase green plantlet productivity to about 7.8%. Therefore, increasing Fe2+in induction media properly could increase anther culture efficiency of early Japonica rice in cold region.展开更多
The immunohistochemical localization of IAA and the comparison of their relative levels were carried out for the first time in the anthers of Nongken 58S and its wild type Nongken 58 (Oryza sativa subsp. japonica) af...The immunohistochemical localization of IAA and the comparison of their relative levels were carried out for the first time in the anthers of Nongken 58S and its wild type Nongken 58 (Oryza sativa subsp. japonica) after long_day and short_day treatments. The distribution of free_IAA in anthers and its dynamic variation could be reflected by this method. The results showed that the IAA level in the anthers of Nongken 58S after long_day treatment was much lower than that in short_day_treated Nongken 58S and those in wild type Nongken 58 in five stages from pistil and stamen primordia formation to late uninucleate stage. The possible reasons for IAA deficiency in Nongken 58S_LD anthers and its relationship with fertility alteration were also discussed.展开更多
Effects of four culture media including MS, N6, C17 and K on wheat anther callus induction in vitro culture were studied. The results showed that the callus in- duction rate of four kinds of culture medium was in the ...Effects of four culture media including MS, N6, C17 and K on wheat anther callus induction in vitro culture were studied. The results showed that the callus in- duction rate of four kinds of culture medium was in the order of K〉C17〉N6〉MS.展开更多
To investigate the culture technique in anther of Chinese wolfberry,we optimized the culture medium(including hormone combination)and culture conditions.The results showed that calluses were induced from all the six t...To investigate the culture technique in anther of Chinese wolfberry,we optimized the culture medium(including hormone combination)and culture conditions.The results showed that calluses were induced from all the six tested Chinese wolfberry materials,but the induction rate of callus varied toward the materials with different genotypes.When the experimental materials were cultured on medium appended with 2,4-D 1.0 mg/L and KT 1.0 mg/L under dark,the callus induction rate reached 20.0 % in this study,and this hormone combination should be the optimum for anther culture of Chinese wolfberry.With MS appended with 6-BA 0.5 mg/L and NAA 0.1 mg/L as differentiation medium and that appended with NAA 0.1 mg/L,the plants could be yielded in 20 days.展开更多
[Objective]In order to increase anther culture efficiency of pepper.[Method]MS culture media and Bolajiaohong were used in this experiment to study the influences of carbon sources and concentrations on anther callus ...[Objective]In order to increase anther culture efficiency of pepper.[Method]MS culture media and Bolajiaohong were used in this experiment to study the influences of carbon sources and concentrations on anther callus induction of pepper.Jiayu was taken as a material to study influences of plant growth regulators and concentrations on anther callus induction of pepper according to L16（4^5） orthogonal design.[Result]The average callus and embryoid induction rates of maltose at all concentrations were higher than these of sucrose but the difference was not significant.Taking maltose or sucrose as a carbon source,3% to 6% concentration was good for increasing induction frequencies of calli and embryoids.However,If the concentration was over 6%,the induction rates were declined dramatically with the increase of sugar concentration.The influences of growth regulators on induction rate of calli were listed as 2,4-D﹥ZT﹥NAA﹥KT﹥6-BA;the influences on induction rates of embryoids were listed as 2,4-D﹥NAA﹥ZT﹥KT﹥6-BA.The 2,4-D,ZT,NAA and KT had signficant or extremely significant influences on induction rates of calli and embryoids.2,4-D,ZT at 1.0 mg/L and NNA,KT at 0.5 mg/L had the best effects.The influences of ZT on calli and embryoids were better than those of KT and 6-BA.1.0 mg/L 2,4-D ＋1.0 mg/L ZT ＋0.5 mg/L KT ＋0.5 mg/L 6-BA was the best regulator combination for induction culture of Jiayu anther.[Conclusion]The experiment provided research basis for anther culture of pepper.展开更多
[Objective] The aim was to investigate the application of plant growth regulator thidiazuron（TDZ）in strawberry anther culture.[Method]Different combinations of TDZ and naphthalene acetic acid（NAA）auxine with diffe...[Objective] The aim was to investigate the application of plant growth regulator thidiazuron（TDZ）in strawberry anther culture.[Method]Different combinations of TDZ and naphthalene acetic acid（NAA）auxine with different concentrations were used for induction culture,and then its effect on callus induction and plant regeneration were observed.[Result] The combination of 1.0 mg/L TDZ＋0.5 mg/LNAA was effective to accelerating the differentiation and regeneration of strawberries,and the differentiation ratio was as high as 75%.However,TDZ showed insignificant effect on strawberries anther induction.[Conclusion]This research had provided reference for large-scale production of strawberry through anther culture.展开更多
Objective] This study was conducted to investigate the effects of different hormone combinations and concentrations on cal us induction and pIant regeneration of F1 hybrid of Solanum torvum SW. ＆#215; Solanum melonge...Objective] This study was conducted to investigate the effects of different hormone combinations and concentrations on cal us induction and pIant regeneration of F1 hybrid of Solanum torvum SW. ＆#215; Solanum melongena L. [Method] Media suppIemented with different combinations and concentrations of hormones were pre-pared for anther cal us induction, proIiferation, differentiation and regeneration. [Re-sult] MS＋1.0 m/L 2, 4-D＋0.5 mg/L 6-BA was the optimum medium for anther cal us induction. MS＋1.0 mg/L KT＋0.1 mg/L NAA was the optimum medium for cal us proIiferation. MS＋2.0 mg/L 6-BA＋0.3 mg/L NAA was the optimum medium for in-duction of adventitious buds. MS＋0.3 mg/L IAA was the optimum medium for root-ing of adventitious buds. The highest rooting rate was onIy 37.5 because induction of rooting was a difficuIt work. [Conclusion] The resuIts wiI provide references for outcross fertiIity and genetic improvement of eggpIant.展开更多
Microsporogenesis and male gametogenesis are essential for the alternating life cycle of flowering plants between diploid sporophyte and haploid gametophyte generations. Rice （Oryza sativa） is the world＇s major sta...Microsporogenesis and male gametogenesis are essential for the alternating life cycle of flowering plants between diploid sporophyte and haploid gametophyte generations. Rice （Oryza sativa） is the world＇s major staple food, and manipulation of pollen fertility is particularly important for the demands to increase rice grain yield. Towards a better understanding of the mechanisms controlling rice male reproductive development, we describe here the cytological changes of anther development through 14 stages, including cell division, differentiation and degeneration of somatic tissues consisting of four concentric cell layers surrounding and supporting reproductive cells as they form mature pollen grains through meiosis and mitosis. Furthermore, we compare the morphological difference of anthers and pollen grains in both monocot rice and eudicot Arabidopsis thaliana. Additionally, we describe the key genes identified to date critical for rice anther development and pollen formation.展开更多
To understand the molecular mechanism of male reproductive development in the model crop rice,we isolated a complete male sterile mutant post-meiotic deficient anther1 （pda1） from a γ-ray-treated rice mutant librar...To understand the molecular mechanism of male reproductive development in the model crop rice,we isolated a complete male sterile mutant post-meiotic deficient anther1 （pda1） from a γ-ray-treated rice mutant library.Genetic analysis revealed that the pda1 mutant was controlled by a recessive nucleus gene.The pda1 mutant anther seemed smaller with white appearance.Histological analysis demonstrated that the pda1 mutant anther undergoes normal early tapetum development without obvious altered meiosis.However,the pda1 mutant displayed obvious defects in postmeiotic tapetal development,abnormal degeneration occurred in the tapetal cells at stage 9 of anther development.Also we observed abnormal lipidic Ubisch bodies from the tapetal layer of the pda1 mutant,causing no obvious pollen exine formation.RT-PCR analysis indicated that the expression of genes involved in anther development including GAMYB,OsC4 and Wax-deficient anther1 （WDA1） was greatly reduced in the pda1 mutant anther.Using map-based cloning approach,the PDA1 gene was finely mapped between two markers HLF610 and HLF627 on chromosome 6 using 3,883 individuals of F2 population.The physical distance between HLF610 and HLF627 was about 194 kb.This work suggests that PDA1 is required for post-meiotic tapetal development and pollen/microspore formation in rice.展开更多
Two rice maintaining lines with different drought tolerances, viz., Jin 23B （tolerant） and Zhenshan 97B （sensitive）, were used to study the oxidative stress and soluble sugar in rice anthers and pollen viability u...Two rice maintaining lines with different drought tolerances, viz., Jin 23B （tolerant） and Zhenshan 97B （sensitive）, were used to study the oxidative stress and soluble sugar in rice anthers and pollen viability under drought stress during flowering stage. Higher antioxidant enzyme activities and lower malonaldehyde （MDA） content in rice anthers were observed in Jin 23B than in Zhenshan 97B under drought stress. Further, a great increase in the content of soluble sugar in rice anthers of Jin 23B was observed across the whole drought exposure, while Zhenshan 97B showed significant decrease in soluble sugar during 9-12 d after drought stress （DADS）. Accordingly, a marked decline of pollen fertility and activity, pollen numbers in an anther and pollen numbers on a stigma was observed in Zhenshan 97B, whereas little difference was found in Jin 23B. Thus, we suggest that pollen abortion caused by drought stress may be related with the reciprocity between oxidative stress and soluble sugar content in rice anthers.展开更多
MADS-box gene family encodes a large number and variety of transcription regulators in plants. In this study, a cDNA, GhMADS9, encoding a typical MADS protein with 230 amino acids was isolated from cotton flower cDNA ...MADS-box gene family encodes a large number and variety of transcription regulators in plants. In this study, a cDNA, GhMADS9, encoding a typical MADS protein with 230 amino acids was isolated from cotton flower cDNA library. Subsequently, a 1,623 bp genomic DNA fragment of GhMADS9 gene was isolated in cotton by PCR. Compared with its cDNA sequence, six introns were found in GhMADS9 gene. Fluorescent microscopy indicated that GhMADS9 protein localized in the nucleus. Transactivation activity assay in yeast cells revealed that GhMADS9 protein did not show transcriptional activation. Quantitative RT-PCR analysis showed that GhMADS9 was specially expressed in cotton anthers. Further in situ hybridization analysis demonstrated that strong expression of GhMADS9 gene was detected in developing pollens, but no or weak signals were found in the other anther tissues. Furthermore, GhMADS9 expression was dramatically up-regulated in anthers with abscisic acid (ABA) treatment, whereas its activity was down-regulated when treated by gibberellin (GA3). Collectively, our results suggest that GhMADS9 is a transcription factor and might be involved in cotton anther/pollen development and in response to ABA and GA3 signaling.展开更多
The ＇double T-DNA＇ binary vector p13HSR which harbored two independent T-DNAs, containing hygromycin phosphotransferase gene （hpf） in one T-DNA region and three target genes （hLF, SB401, RZ10） in another T-DNA r...The ＇double T-DNA＇ binary vector p13HSR which harbored two independent T-DNAs, containing hygromycin phosphotransferase gene （hpf） in one T-DNA region and three target genes （hLF, SB401, RZ10） in another T-DNA region, was used to generate selectable marker-free transgenic rice by Agrobacterium-mediated transformation. The regenerated plants with both the three target genes and the selectable marker gene hpt were selected for anther culture. RT-PCR analysis indicated that target genes were inserted in rice genomic DNA and successfully transcribed. It took only one year to obtain double haploid selectable marker-free transgenic plants containing the three target genes with co-transformation followed by anther culture technique, and the efficiency was 12.2%. It was also noted that one or two target genes derived from the binary vector were lost in some transgenic rice plants.展开更多
The anther cuticle and pollen exine play a critical role in male gametophyte development. The sporopollenin precursors and cuticular lipid monomers are transported to the surface of the microspores and the epidermis b...The anther cuticle and pollen exine play a critical role in male gametophyte development. The sporopollenin precursors and cuticular lipid monomers are transported to the surface of the microspores and the epidermis by lipid transport proteins(LTPs) and ATP-binding cassette G(ABCG) transporters for the formation of the pollen wall and anther cuticle, respectively. However, the function of ABCG transporters in maize anther development is unclear. Here, we cloned the MS2 gene from the maize male sterile2 mutant using map-based cloning and determined that it encodes an ABCG transporter. MS2 protein was experimentally confirmed to be located on the cell membrane. The quantitative real-time PCR(qRT-PCR)results showed that MS2 was ubiquitously expressed in all vegetative and reproductive tissues, whereas a high transcriptional level of MS2 was observed in anthers, especially at the young microspore stage. Gas chromatography-mass spectrometry(GC–MS) analysis showed decreased accumulation of cutin and wax components in ms2 anthers, indicating that MS2 plays a role in the transport of lipid molecules to anther cuticle and pollen exine. To our knowledge, MS2 is the first reported ABCG transporter gene that participates in anther development in maize.展开更多
基金supported by the Science and Technology Innovation Program of Fujian Agriculture and Forestry University,China(Grant No.KFB22045)the General Program of Natural Science Foundation Fujian,China(Grant No.2023J01460).
文摘The doubled haploid(DH)technique accelerates homozygosity by inducing chromosome doubling in haploid embryos derived from hybrid plants.This approach offers significant advantages over conventional rice breeding methods by shortening the breeding cycle and enabling rapid development of pure homozygous lines.Anther culture(AC)has been established as an efficient and successful method for producing DH plants via androgenesis in rice.However,despite its success in japonica rice.
基金the National Natural Science Foundation of China(32170271,32470277)the Project of Sanya Yazhou Bay Science and Technology City(SCKJ-JYRC-2023-52)the Natural Science Foundation of Henan Province(252300421076,222300420024).
文摘Carotenoids are lipophilic isoprenoid pigments with essential roles in plants.While the cultivated allotetraploid cottons exhibit distinct mature anther coloration—yellow in Gossypium barbadense versus predominantly white in G.hirsutum—the genetic basis of this divergence remains unclear.The purpose of this study was to identify the genetic basis of anther-color variation in cotton(Gossypium)species.We firstly identified carotenoids as the primary pigments underlying yellow-anthers coloration.Comparative transcriptomics of anthers revealed that the carotenoid biosynthesis gene GbPSY4 was expressed as a key regulator in G.barbadense.Functional validation via tissue-specific expression,subcellular localization,in vivo enzymatic assays,and virus-induced gene silencing confirmed its role in carotenoid biosynthesis and yellow pigmentation.Genome-wide association studies in a G.hirsutum population revealed GhPSY4_At,an ortholog of GbPSY4,as the causal gene of anther-color variation.We conclude that PSY4-regulated carotenoid biosynthesis governs yellow pigmentation.Furthermore,a finding that G.hirsutum accessions with yellow anthers showed greater pollen viability under high-temperature stress than those with white anthers suggests that the same pathway that governs yellow pigmentation influences heat tolerance.PSY4 is a promising target for breeding stress-tolerant cotton varieties.
基金supported by the Science and Technology Major Program of Bingtuan,China (2023AA008)the National Natural Science Foundation of China (31960369)+1 种基金the Bingtuan Science and Technology Program,China (2025DA001)the Henan Provincial Science and Technology Research Project,China (222102110200)。
文摘Recent studies have shown that lipid metabolism is a key factor affecting anther development and male fertility.However,how plants regulating the metabolic balance of multiple lipids to ensure proper anther development and male fertility remains unclear.Analyzing lipid molecules related to anther fertility and genes responsible for their biosynthesis is crucial for understanding the physiological significance of lipid metabolism in crop fertility.In this study,we compared the transcriptome and the composition and content of lipids in anthers of two upland cotton(Gossypium hirsutum) materials,Shida 98(WT) and its nearly-isogenic male sterile line Shida 98A(MS).Transcriptomics analysis identified many differentially expressed genes(DEGs) between the two materials,with the genes of the alpha-linolenic acid metabolism pathway being the most significantly associated with the male sterility phenotype.Investigations on lipids revealed that the MS anthers over-accumulated free fatty acids(FFAs),phosphatidic acid(PA),mono-and di-galactosyldiacylglycerol(MGDG and DGDG),and had a decreased content of triacylglycerol(TAG),which was closely related to the abnormal metabolism of alpha-linolenic acid(C18:3);therefore,the major lipids containing C18:3-acyl chains,such as PA,MGDG,DGDG,and TAG,are proposed to play a major role in cotton anther development.We also showed that an excessive level of MGDG and DGDG caused jasmonic acid(JA) overaccumulation in MS anthers,which in turn inhibited the expression of GhFAD3 and consequently reduced the C18:3 content,presumably via a feedback regulation mechanism,ultimately affecting plant fertility.Together,our results revealed the importance of a balanced lipid metabolism in regulating the development of cotton anther and pollen and consequently male fertility.
基金financially supported in part by grants from Biological Breeding-National Science and Technology Major Project (2023ZD04040)Jiangsu Key Research and Development Program (BE2022384)Collaborative Innovation Center for Modern Crop Production co-sponsored by Province and Ministry (CICMCP) (No. 10)
文摘Carotenoids are natural pigments that are widely distributed in the flowers,fruits,and seeds of many plant species.These compounds not only endow diverse colors but also exhibit antioxidant,immune-modulatory,anti-aging,and photoprotective properties.Although carotenoid metabolism has been studied extensively in microbial and plant science,the genetic mechanisms underlying carotenoid metabolism in cotton remain underexplored.Here,we isolated gene GbDYA that regulates a dark-yellow anther color by map-based cloning using a BC1F1 population derived from a cross of Gossypium barbadense acc.Hai7124 with dark-yellow anthers and G.hirsutum acc.TM-1 with light-yellow anthers backcrossed with TM-1.GbDYA encodes phytoene synthase,a key rate-limiting enzyme in the carotenoid biosynthesis pathway.A long terminal repeat retrotransposon in the first exon of GhDYA(an ortholog of GbDYA in G.hirsutum acc.TM-1)caused loss of function and led to the light-yellow anther color.GbDYA is predominantly expressed in the early stages of anther development.Transcriptome,RT-qPCR and KEGG enrichment analyses revealed that GbDYA influences the synthesis and accumulation of carotenoids in anthers by modulating expression of key genes in the carotenoid biosynthesis pathway.Integrated transcriptomic and metabolomic analyses indicated that the accumulation of lutein,violaxanthin,antherxanthin,cryptoxanthin,zeaxanthin,andβ-carotene contributed to yellow coloration of anthers.Dual-luciferase and yeast one-hybrid assays confirmed that transcription factor GbMYB105(GB_A11G3511)binds to the promoter of GbDYA and activates its expression.High-temperature stress treatment indicated that carotenoids accumulation in anthers enhances pollen antioxidant activity.This study unravels the role of GbDYA in conferring the anther coloration,and provides the potential utilization by modulating accumulation of carotenoids in anthers to enhance pollen viability in high-temperature tolerance breeding in cotton.
基金supported by the Excellent Young Scientists Fund(Category B)(32422063)the National Key Research and Development Program of China(2022YFF1003500)the Zhengzhou University Qiushi Postdoctoral Research Funding Program.For open access,the authors have applied for a Creative Commons Attribution(CC BY)license for any Author Accepted Manuscript version arising from this submission.
文摘Anther is a key male reproductive organ that is essential for the plant life cycle,from the sporophyte to the gametophyte generation.To explore the isoform-level transcriptional landscape of developing anthers in maize(Zea mays L.),we analyzed Iso-Seq data from anthers collected at 10 developmental stages,together with strand-specific RNA-seq,CAGE-seq,and PAS-seq data.Of the 152,026 high-confidence full-length isoforms identified,68.8%have not been described;these include 22,365 isoforms that originate from previously unannotated loci and 82,167 novel isoforms that originate from annotated protein-coding genes.Using our newly developed strategy to detect dynamic expression patterns of isoforms,we identify 13,899 differentially variable regions(DVRs);surprisingly,1275 genes contain more than two DVRs,revealing highly efficient utilization of limited genic regions.We identify 7876 long non-coding RNAs(lncRNAs)from 4098 loci,most of which were preferentially expressed during cell differentiation and meiosis.We also detected 371 long-range interactions involving intergenic lncRNAs(lincRNAs);interestingly,243 were lincRNA-gene ones,and the interacting genes were highly expressed in anthers,suggesting that many potential lncRNA regulators of key genes are required for anther development.This study provides valuable resources and fundamental information for studying the essential transcripts of key genes during anther development.
文摘Water scarcity is an escalating global challenge that severely threatens productivity and reproductive success in crops,particularly in drought-sensitive species such as Capsicum annuum L.Although deficit irrigation strategies are widely recommended to enhance water use efficiency,knowledge remains limited regarding their interactions with soil amendments such as biochar and the consequent impacts on reproductive traits.This study aimed to evaluate the combined effects of deficit irrigation strategies and biochar application on pollen viability and morphology in Capsicum annuum.The experiment was conducted under full,partial,and deficit irrigation regimes with and without biochar treatment,following a randomized block design.The primary parameters examined were pollen viability(viable,semi-viable,and non-viable rates),anther width and length,and pollen width and length.Microscopic measurements and statistical analyses(p≤0.05)revealed significant effects of both irrigation regimes and biochar applications.Under deficit irrigation,viable,semi-viable,and non-viable pollen rates were 29.84%,32.95%,and 37.21%,respectively,whereas the highest viable pollen rate was observed under full irrigation.In partial irrigation,viable pollen accounted for 31.67%,semi-viable for 38.81%,and non-viable for 29.49%.In plots treated with biochar under partial irrigation,anther width(1700.89μm),anther length(3805.34μm),pollen width(26.93μm),and pollen length(37.42μm)reached the highest values,while the lowest values were recorded in deficit irrigation plots without biochar.These findings emphasize the importance of integrating biochar into irrigation management to mitigate the adverse effects of water stress on pollen development.Nevertheless,further research is needed to clarify the long-term implications of these practices for reproductive success and agricultural sustainability.
基金Supported by the National 863 Project of Tenth-five Year Plan(2001AA2411042004AA241104)+1 种基金Key Breeding Project of Sichuan Province and(200107001-16-01)Key Quality Project of Sichuan Province(200107001-1-7-4)~~
文摘Some influential factors of anther culture were studied preliminarily by conducting anther culture of the restorers of new cytoplasmic male sterile （NER）. Several results were obtain from this experiment and they were listed as follow：① MS cultrure medium with such hormones as 2,4-D 2 mg/L,6-BA 0.5 mg/L, NAA 0.5 mg/L was the best suitable for callus induction of NER. ②The difference of induction rate was significantly different between different plant age groups. From the 110th day to 141th day,the induction rate was increased with the increase of age and the difference of induction rate reached 0.01 significant difference level. The induction rate reached the highest value in the 141th day then it declined gradually. ③The combined use of 2, 4-D and 6-BA with proper increase of 2,4-D was good for inducing callus. ④The green plantlet induction rate of NER was increased when the concentration of 6-BA increased from 2 mg/L to 4 mg/L. Adding ZT from 0.5 mg/L to 2 mg/L. 6-BA would led 2.47% increase of green plantlet olantlet induction rate.
基金Supported by National Key Technology R&D Program(2006BAD01A01-6)~~
文摘In this study, through vitro culturing anthers of 7 F1 progenies of early Japonica rice in cold region on medium with different Fe2+ contents, it was found that Fe2+ content generated greater impacts on the induction rate and green plantlet differentiation. The result demonstrated that if Fe2+ increased from 32 to 40 mg/kg, the induction rate of early Japonica rice anther culture in N6 culture media was more then 1.4 times higher than that in N6 culture media containing 5.6 mg/kg Fe2+. In this concentration range, the induction rate increased with the increase of Fe2+ content, while if the concentration was over this concentration range, the induction rate decreased with the increase of Fe2+, showing single peak distribution. When the Fe2+ was 40 mg/kg in differentiation medium, the differentiation rate decreased dramatically. The green plantlet differentiations of callus which were induced on culture media containing 32-40 mg/kg Fe2+ were different, when they were cultured on MS culture media, and 85.7% materials could increase green plantlet productivity to about 7.8%. Therefore, increasing Fe2+in induction media properly could increase anther culture efficiency of early Japonica rice in cold region.
文摘The immunohistochemical localization of IAA and the comparison of their relative levels were carried out for the first time in the anthers of Nongken 58S and its wild type Nongken 58 (Oryza sativa subsp. japonica) after long_day and short_day treatments. The distribution of free_IAA in anthers and its dynamic variation could be reflected by this method. The results showed that the IAA level in the anthers of Nongken 58S after long_day treatment was much lower than that in short_day_treated Nongken 58S and those in wild type Nongken 58 in five stages from pistil and stamen primordia formation to late uninucleate stage. The possible reasons for IAA deficiency in Nongken 58S_LD anthers and its relationship with fertility alteration were also discussed.
基金Supported by the National Natural Science Foundation of China(31071413)the Key Scientific and Technological Project of Henan Province(122102110189)+2 种基金the Natural Science Foundation of Henan Province(2011B210002)the Project for the Youth of Henan Institute of Education(20090103)the Key Research Project of Institution of Higher Education(15A210020)~~
文摘Effects of four culture media including MS, N6, C17 and K on wheat anther callus induction in vitro culture were studied. The results showed that the callus in- duction rate of four kinds of culture medium was in the order of K〉C17〉N6〉MS.
基金Supported by Projects of Sci-Tech Achievements Transformation in Chi-na(2006GBZG300311)National Natural Science Foundation of China(30760127)~~
文摘To investigate the culture technique in anther of Chinese wolfberry,we optimized the culture medium(including hormone combination)and culture conditions.The results showed that calluses were induced from all the six tested Chinese wolfberry materials,but the induction rate of callus varied toward the materials with different genotypes.When the experimental materials were cultured on medium appended with 2,4-D 1.0 mg/L and KT 1.0 mg/L under dark,the callus induction rate reached 20.0 % in this study,and this hormone combination should be the optimum for anther culture of Chinese wolfberry.With MS appended with 6-BA 0.5 mg/L and NAA 0.1 mg/L as differentiation medium and that appended with NAA 0.1 mg/L,the plants could be yielded in 20 days.
文摘[Objective]In order to increase anther culture efficiency of pepper.[Method]MS culture media and Bolajiaohong were used in this experiment to study the influences of carbon sources and concentrations on anther callus induction of pepper.Jiayu was taken as a material to study influences of plant growth regulators and concentrations on anther callus induction of pepper according to L16（4^5） orthogonal design.[Result]The average callus and embryoid induction rates of maltose at all concentrations were higher than these of sucrose but the difference was not significant.Taking maltose or sucrose as a carbon source,3% to 6% concentration was good for increasing induction frequencies of calli and embryoids.However,If the concentration was over 6%,the induction rates were declined dramatically with the increase of sugar concentration.The influences of growth regulators on induction rate of calli were listed as 2,4-D﹥ZT﹥NAA﹥KT﹥6-BA;the influences on induction rates of embryoids were listed as 2,4-D﹥NAA﹥ZT﹥KT﹥6-BA.The 2,4-D,ZT,NAA and KT had signficant or extremely significant influences on induction rates of calli and embryoids.2,4-D,ZT at 1.0 mg/L and NNA,KT at 0.5 mg/L had the best effects.The influences of ZT on calli and embryoids were better than those of KT and 6-BA.1.0 mg/L 2,4-D ＋1.0 mg/L ZT ＋0.5 mg/L KT ＋0.5 mg/L 6-BA was the best regulator combination for induction culture of Jiayu anther.[Conclusion]The experiment provided research basis for anther culture of pepper.
基金Supported by Science and Technology Research Projects in Ningxia Hui Autonomous Region(KGZ-16-07-02)Opening project in National Engineering Laboratory of Tree Breeding in Beijing Forestry University~~
文摘[Objective] The aim was to investigate the application of plant growth regulator thidiazuron（TDZ）in strawberry anther culture.[Method]Different combinations of TDZ and naphthalene acetic acid（NAA）auxine with different concentrations were used for induction culture,and then its effect on callus induction and plant regeneration were observed.[Result] The combination of 1.0 mg/L TDZ＋0.5 mg/LNAA was effective to accelerating the differentiation and regeneration of strawberries,and the differentiation ratio was as high as 75%.However,TDZ showed insignificant effect on strawberries anther induction.[Conclusion]This research had provided reference for large-scale production of strawberry through anther culture.
基金Supported by National Science and Technology Program for Rural Development of China during the 12thFive-Year Plan Period(2012AA100103007)National Agricultural Science and Technology Achievement Transformation Fund(2013GB2E100381)+1 种基金Earmarked Fund for China Agriculture Research System(nycytxgxcxtd-03-10)Science and Technology Program of Guangxi(14123006-35,14123004-3-5)~~
文摘Objective] This study was conducted to investigate the effects of different hormone combinations and concentrations on cal us induction and pIant regeneration of F1 hybrid of Solanum torvum SW. ＆#215; Solanum melongena L. [Method] Media suppIemented with different combinations and concentrations of hormones were pre-pared for anther cal us induction, proIiferation, differentiation and regeneration. [Re-sult] MS＋1.0 m/L 2, 4-D＋0.5 mg/L 6-BA was the optimum medium for anther cal us induction. MS＋1.0 mg/L KT＋0.1 mg/L NAA was the optimum medium for cal us proIiferation. MS＋2.0 mg/L 6-BA＋0.3 mg/L NAA was the optimum medium for in-duction of adventitious buds. MS＋0.3 mg/L IAA was the optimum medium for root-ing of adventitious buds. The highest rooting rate was onIy 37.5 because induction of rooting was a difficuIt work. [Conclusion] The resuIts wiI provide references for outcross fertiIity and genetic improvement of eggpIant.
基金supported by the funds from the National Basic Research Program of China(Nos.2009CB941500 and 2007CB108700)the National Natural Science Foundation of China(No.30725022)+1 种基金the Chinese Transgenic Project(No. 2009ZX08009-108B)the National 863 High-Tech Project (No.2011AA10A101)
文摘Microsporogenesis and male gametogenesis are essential for the alternating life cycle of flowering plants between diploid sporophyte and haploid gametophyte generations. Rice （Oryza sativa） is the world＇s major staple food, and manipulation of pollen fertility is particularly important for the demands to increase rice grain yield. Towards a better understanding of the mechanisms controlling rice male reproductive development, we describe here the cytological changes of anther development through 14 stages, including cell division, differentiation and degeneration of somatic tissues consisting of four concentric cell layers surrounding and supporting reproductive cells as they form mature pollen grains through meiosis and mitosis. Furthermore, we compare the morphological difference of anthers and pollen grains in both monocot rice and eudicot Arabidopsis thaliana. Additionally, we describe the key genes identified to date critical for rice anther development and pollen formation.
基金supported by the Funds from the National Key Basic Research Developments Program of the Ministry of Science and Technology, China (No. 2009CB941500)Shanghai Leading Academic Discipline Project (No. B205)
文摘To understand the molecular mechanism of male reproductive development in the model crop rice,we isolated a complete male sterile mutant post-meiotic deficient anther1 （pda1） from a γ-ray-treated rice mutant library.Genetic analysis revealed that the pda1 mutant was controlled by a recessive nucleus gene.The pda1 mutant anther seemed smaller with white appearance.Histological analysis demonstrated that the pda1 mutant anther undergoes normal early tapetum development without obvious altered meiosis.However,the pda1 mutant displayed obvious defects in postmeiotic tapetal development,abnormal degeneration occurred in the tapetal cells at stage 9 of anther development.Also we observed abnormal lipidic Ubisch bodies from the tapetal layer of the pda1 mutant,causing no obvious pollen exine formation.RT-PCR analysis indicated that the expression of genes involved in anther development including GAMYB,OsC4 and Wax-deficient anther1 （WDA1） was greatly reduced in the pda1 mutant anther.Using map-based cloning approach,the PDA1 gene was finely mapped between two markers HLF610 and HLF627 on chromosome 6 using 3,883 individuals of F2 population.The physical distance between HLF610 and HLF627 was about 194 kb.This work suggests that PDA1 is required for post-meiotic tapetal development and pollen/microspore formation in rice.
基金supported by the National Natural Science Foundation of China (30871473)the project from the China National Rice Research Institute (2009RG0035)+1 种基金the projects from Local Government of Zhejiang Province, China (2008C22073, 2009C32048, and SN200806)the Zhejiang Provincial Natural Science Foundation, China (Y3100060 and Y3100090)
文摘Two rice maintaining lines with different drought tolerances, viz., Jin 23B （tolerant） and Zhenshan 97B （sensitive）, were used to study the oxidative stress and soluble sugar in rice anthers and pollen viability under drought stress during flowering stage. Higher antioxidant enzyme activities and lower malonaldehyde （MDA） content in rice anthers were observed in Jin 23B than in Zhenshan 97B under drought stress. Further, a great increase in the content of soluble sugar in rice anthers of Jin 23B was observed across the whole drought exposure, while Zhenshan 97B showed significant decrease in soluble sugar during 9-12 d after drought stress （DADS）. Accordingly, a marked decline of pollen fertility and activity, pollen numbers in an anther and pollen numbers on a stigma was observed in Zhenshan 97B, whereas little difference was found in Jin 23B. Thus, we suggest that pollen abortion caused by drought stress may be related with the reciprocity between oxidative stress and soluble sugar content in rice anthers.
基金supported by the National Natural Sciences Foundation of China (No. 30871317)the project from Ministry of Agriculture of China for transgenic research (No. 2009ZX08009-117B)
文摘MADS-box gene family encodes a large number and variety of transcription regulators in plants. In this study, a cDNA, GhMADS9, encoding a typical MADS protein with 230 amino acids was isolated from cotton flower cDNA library. Subsequently, a 1,623 bp genomic DNA fragment of GhMADS9 gene was isolated in cotton by PCR. Compared with its cDNA sequence, six introns were found in GhMADS9 gene. Fluorescent microscopy indicated that GhMADS9 protein localized in the nucleus. Transactivation activity assay in yeast cells revealed that GhMADS9 protein did not show transcriptional activation. Quantitative RT-PCR analysis showed that GhMADS9 was specially expressed in cotton anthers. Further in situ hybridization analysis demonstrated that strong expression of GhMADS9 gene was detected in developing pollens, but no or weak signals were found in the other anther tissues. Furthermore, GhMADS9 expression was dramatically up-regulated in anthers with abscisic acid (ABA) treatment, whereas its activity was down-regulated when treated by gibberellin (GA3). Collectively, our results suggest that GhMADS9 is a transcription factor and might be involved in cotton anther/pollen development and in response to ABA and GA3 signaling.
文摘The ＇double T-DNA＇ binary vector p13HSR which harbored two independent T-DNAs, containing hygromycin phosphotransferase gene （hpf） in one T-DNA region and three target genes （hLF, SB401, RZ10） in another T-DNA region, was used to generate selectable marker-free transgenic rice by Agrobacterium-mediated transformation. The regenerated plants with both the three target genes and the selectable marker gene hpt were selected for anther culture. RT-PCR analysis indicated that target genes were inserted in rice genomic DNA and successfully transcribed. It took only one year to obtain double haploid selectable marker-free transgenic plants containing the three target genes with co-transformation followed by anther culture technique, and the efficiency was 12.2%. It was also noted that one or two target genes derived from the binary vector were lost in some transgenic rice plants.
基金supported by the Fundamental Research Funds for Central Non-Profit of Institute of Crop Sciences, Chinese Academy of Agricultural Sciences (S2018QY07)National Major Project for Transgenic Organism Breeding (2016ZX08010-004)。
文摘The anther cuticle and pollen exine play a critical role in male gametophyte development. The sporopollenin precursors and cuticular lipid monomers are transported to the surface of the microspores and the epidermis by lipid transport proteins(LTPs) and ATP-binding cassette G(ABCG) transporters for the formation of the pollen wall and anther cuticle, respectively. However, the function of ABCG transporters in maize anther development is unclear. Here, we cloned the MS2 gene from the maize male sterile2 mutant using map-based cloning and determined that it encodes an ABCG transporter. MS2 protein was experimentally confirmed to be located on the cell membrane. The quantitative real-time PCR(qRT-PCR)results showed that MS2 was ubiquitously expressed in all vegetative and reproductive tissues, whereas a high transcriptional level of MS2 was observed in anthers, especially at the young microspore stage. Gas chromatography-mass spectrometry(GC–MS) analysis showed decreased accumulation of cutin and wax components in ms2 anthers, indicating that MS2 plays a role in the transport of lipid molecules to anther cuticle and pollen exine. To our knowledge, MS2 is the first reported ABCG transporter gene that participates in anther development in maize.
