Lilies are widely cultivated for cut flowers,but their large anthers carry a considerable amount of colored pollen that is dispersed easily.Studying the molecular mechanism of anther development and dehiscence could h...Lilies are widely cultivated for cut flowers,but their large anthers carry a considerable amount of colored pollen that is dispersed easily.Studying the molecular mechanism of anther development and dehiscence could help solve this problem.LoMYB21,encoding a putative R2R3v-myb avian myeloblastosis viral oncogene homolog(MYB)transcription factor,was identified from oriental lilies(Lilium‘Siberia’).Real-time quantitative PCR analysis showed that LoMYB21 was mainly expressed in the anther,filament and stigma and had high expression during the late stages of lily anther development.LoMYB21 had transactivation activity and was located in the nucleus through yeast one-hybrid assays and transient expression in Nicotiana benthamiana.Suppression of LoMYB21 by virus-induced gene silencing(VIGS)in Lilium‘Siberia’led to anther indehiscence and reduced the expression of genes related to Jasmonate acid(JA)biosynthesis and signal transduction.Induction of LoMYB21 in DEX::LoMYB21 transgenic Arabidopsis caused procumbent inflorescences that became infertile,accompanied by higher expression of JA biosynthetic and signaling genes.These results demonstrated that JA content and signaling were abnormal in silenced lily and transgenic LoMYB21 Arabidopsis,which affected anther development.Our study indicated that LoMYB21 could regulate lily anther dehiscence through JA biosynthesis and signaling during the late stages of anther development.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
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
Lily(Lilium spp.)is an important horticultural crop,but its use is limited due to serious pollen contamination problems.There are many studies on pollen development in model plants,but few on flower crops such as lili...Lily(Lilium spp.)is an important horticultural crop,but its use is limited due to serious pollen contamination problems.There are many studies on pollen development in model plants,but few on flower crops such as lilies.Gibberellin(GA)is a large class of hormones and plays an important role in plant vegetative growth and reproductive development.GAMYB is a group of the R2R3-MYB family upregulated by gibberellin,and plays an important role in anther development.Here,we isolated a novel GAMYB,named LoMYB65,from lily,which was closely related to the AtMYB65 and AtMYB33 in Arabidopsis.Fluorescence quantitative PCR results showed that LoMYB65 was mainly expressed in lily anthers.LoMYB65 could be activated by 288μmol·L^(-1)GA3treatment and the LoMYB65 protein was located in the nucleus and cytoplasm,and had transactivation in yeast and tobacco leaf cells.The conserved motif within 226 amino acids of the C-terminal of LoMYB65 contributed to its transactivation.Overexpression of LoMYB65 caused dwarf phenotype,unnormal tapetum development,less seeds of siliques in transgenic Arabidopsis plants,the transgenic plants showed partly male sterile.Simultaneously,silencing of LoMYB65 with VIGS(Virus Induced Gene Silencing)in lily anthers caused unnormal pollen development and reduced the pollen amount.Overexpression of LoMYB65 in Arabidopsis and silencing of LoMYB65 in lily resulted in decreased pollen counts,so we speculate that LoMYB65 may be dose-dependent.Overall,these findings suggest that LoMYB65 may play an important role in anther development and pollen formation in lily.LoMYB65 may provide a useful candidate gene for pollenless breeding of lily.展开更多
Anther dehiscence controls optimal interaction between pollen and stigma,thereby determining the successful sexual reproduction.The regulators or mechanisms of this process remain elusive.Here,two CRISPR/Cas9 mutants ...Anther dehiscence controls optimal interaction between pollen and stigma,thereby determining the successful sexual reproduction.The regulators or mechanisms of this process remain elusive.Here,two CRISPR/Cas9 mutants of a rice exocyst subunit gene SEC3A,sec3a-1 and sec3a-2,showed anther indehiscence at anthesis and male sterility at maturity.Pollen viability and germination in the mutants were partly defective,whereas their female gametes undergone a normal development.Hybrid or self-pollinated seeds could be produced by artificial pollination,suggesting potential use of a weak sec3a mutant as a female line during hybrid breeding.SEC3A is widely expressed in various tissues,including anther walls.Further results showed an excessive IAA accumulation and no endothecium lignification in sec3a-1/2 anthers.Our findings suggest that SEC3A appears to regulate anther dehiscence by modulating auxin signaling,providing insights into regulation of anther dehiscence and function of exocyst in plants.展开更多
The temporal and spatial expression patterns of calmodulin mRNA in the developing tobacco anthers were investigated by in situ RNA hybridization, using digoxigening-labeled anti-RNA probe. Calmodulin mRNA was distribu...The temporal and spatial expression patterns of calmodulin mRNA in the developing tobacco anthers were investigated by in situ RNA hybridization, using digoxigening-labeled anti-RNA probe. Calmodulin mRNA was distributed in various developmental stages of tobacco anthers, but the expression level had temporal and spatial differences distinctly. During early stage of anther development, the expression level of calmodulin mRNA was significantly high, mainly distributed in epidermis, tapetum and transfusion parenchyma cells and so on. Especially, more mRNA was accumulated in the nuclei and chromosomes of microspore mother cells prior to and during meiosis. With the development of anther,mRNA was decreased gradually in the anther wall and pollen. By mature pollen stage, only a stronger positive reaction still existed in the epidermis of anther wall and transfusion parenchyma cells. The results suggest that the temporal and spatial expression of calmodulin mRNA is closely correlated with cell division,展开更多
Rice（Oryza sativa L.） MADS3 transcription factor regulates the homeostasis of reactive oxygen species（ROS）during late anther development,and one MADS3 mutant,mads3-4,has defective anther walls,aborted microspores ...Rice（Oryza sativa L.） MADS3 transcription factor regulates the homeostasis of reactive oxygen species（ROS）during late anther development,and one MADS3 mutant,mads3-4,has defective anther walls,aborted microspores and complete male sterility.Here,we report the untargeted metabolomic analysis of both wild type and mads3-4 mature anthers.Mutation of MADS3 led to an unbalanced redox status and caused oxidative stress that damages lipid,protein,and DNA.To cope with oxidative stress in mads3-4 anthers,soluble sugars were mobilized and carbohydrate metabolism was shifted to amino acid and nucleic acid metabolism to provide substrates for the biosynthesis of antioxidant proteins and the repair of DNA.Mutation of MADS3 also affected other aspects of rice anther development such as secondary metabolites associated with cuticle,cell wall,and auxin metabolism.Many of the discovered metabolic changes in mads3-4 anthers were corroborated with changes of expression levels of corresponding metabolic pathway genes.Altogether,this comparative metabolomic analysis indicated that MADS3 gene affects rice anther development far beyond the ROS homeostasis regulation.展开更多
The regulation of nutrient transportation and transformation in developing anthers is very complex. We analyzed the distribution and features of polysaccharides and lipids in the developing anthers of tomatoes using h...The regulation of nutrient transportation and transformation in developing anthers is very complex. We analyzed the distribution and features of polysaccharides and lipids in the developing anthers of tomatoes using histochemical methods. Some starches appeared in the connective somatic tissue of anthers during the sporogenous cell stage. Before meiosis of the microspore mother cell,a thick polysaccharide callose wall was formed,accompanied by a reduction in the connective tissue starches. During the tetrad stage after meiosis,the polysaccharide material in the anther did not change. At the early microspore stage,the starches in the connective cells again increased,and polysaccharide material appeared in the partial intine of pollen. At the late microspore stage,a large vacuole formed that did not contain lipids or starches,and only the pollen wall contained red polysaccharides. At this stage,the connective somatic cell starch amounts decreased,and the tapetal cells changed shape and degenerated. After microspore division,abundant lipids appeared in the bicellular pollen,and starches accumulated following pollen development. As the anthers matured,many lipids and some starches accumulated in the epidermal cells. Nutrient metabolism within the tomato pollen characteristically accumulated lipids first and then starches,while the mature pollen accumulated starches and lipids simultaneously. This characteristic pattern of nutrient metabolism in tomato pollen shows species specificity among plants.展开更多
Regeneration of haploid plants from wheat pollen cells by the anther culture method was long ago well accomplished by Ouyang et al. Thereafter, it has been used in wheat breeding. Hu Daofen et al. have made contributi...Regeneration of haploid plants from wheat pollen cells by the anther culture method was long ago well accomplished by Ouyang et al. Thereafter, it has been used in wheat breeding. Hu Daofen et al. have made contribution in wheat anther cultrure breeding and a new variety obtained has been widely used in production. Successes展开更多
Doubled haploid(DH)plants have been widely used for breeding and biological research in crops.Pop ulus spp.have been used as model woody plant species for biological research.However,the induction of DH poplar plants ...Doubled haploid(DH)plants have been widely used for breeding and biological research in crops.Pop ulus spp.have been used as model woody plant species for biological research.However,the induction of DH poplar plants is onerous,and limited biological or breeding work has been carried out on DH individuals or populations.In this study,we provide an effective protocol for poplar haploid induction based on an anther culture method.A total of 96 whole DH plant lines were obtained using an F1hybrid of Populus simonii×P.nigra as a donor tree.The phenotypes of the DH population showed exceptionally high variance when compared to those of half-sib progeny of the donor tree.Each DH line displayed distinct features compared to those of the other DH lines or the donor tree.Additionally,some excellent homozygous lines have the potential to be model plants in genetic and breeding studies.展开更多
Callose contributes to many biological processes of higher plants including pollen development,cell plate and vascular tissue formation,as well as regulating the transport function of plasmodesmata.The functions of ca...Callose contributes to many biological processes of higher plants including pollen development,cell plate and vascular tissue formation,as well as regulating the transport function of plasmodesmata.The functions of callose synthase genes in maize have been little studied.We describe a maize male-sterile mutant 39(ms39)characterized by reduced plant height.In this study,we confirmed using CRISPR/Cas9 technology that a mutation in Zm00001d043909(ZmCals12),encoding a callose synthase,is responsible for the male sterility of the ms39 mutant.Compared with male-fertile plants,callose deposition around the dyads and tetrads in ms39 anthers was significantly reduced.Increased cell autophagy observed in ms39 anthers may have been due to the premature programmed cell death of tapetal cells,leading to collapse of the anther wall structure.Disordered glucose metabolism in ms39 may have intensified autophagy in anthers.Evaluation of the ms39 gene on maize heterosis by paired-crossed experiment with 11 maize inbred lines indicated that ms39 can be used for maize hybrid seed production.展开更多
Plant cell elongation depends on well-defined gene regulations,adequate nutrients,and timely cell wall modifications.Anther size is positively correlated with the number and viability of pollen grains,while little is ...Plant cell elongation depends on well-defined gene regulations,adequate nutrients,and timely cell wall modifications.Anther size is positively correlated with the number and viability of pollen grains,while little is known about molecular mechanisms underlying anther cell elongation.Here,we found that properly activated cell elongation regulators at transcriptional levels in loss-of-function ZmMs33 mutant(ms33-6038)anthers failed to promote maize anther elongation.ZmMs33 deficiency disrupted metabolic homeostasis mainly by inhibiting both photosynthesis in anther endothecium and lipid accumulation in anther tapetum.Importantly,ms33-6038 anthers displayed ectopic,premature and excessive secondary cell wall thickening in anther middle layer,which constrained cell elongation structurally and blocked nutrient flows across different anther wall layers.The metabolic disorder was only found in ms33-6038 mutant rather than several representative male-sterility lines at transcriptional and post-translational levels.Collectively,the disordered metabolisms and blocked nutrient flows defeated the activated cell elongation regulators,and finally inhibited anther elongation and growth with a unique‘‘idling effect”in ms33-6038 mutant.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
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
This paper looks back to the development of plant tissue culture in China in the last century. Since 1934, tissue culture studies in China has kept up with the international development in the fields. Progress has bee...This paper looks back to the development of plant tissue culture in China in the last century. Since 1934, tissue culture studies in China has kept up with the international development in the fields. Progress has been made by Chinese in nearly every branches of tissue culture, including in vitro organogenesis, shoot tip culture, anther culture, ovary culture, endosperm culture, protoplast culture as well as mass cell culture. On the basis of reviewing the articles written by Chinese on plant tissue culture, the internationally recognized contributions are specially mentioned. The applications of plant tissue culture to agriculture and industry in China are also introduced.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
基金funded by National Key R&D Program of China(Grant Nos.2020YFD1000402,2018YFD1000400)Chinese Universities Scientific Fund(Grant Nos.2021TC102,2018QC096).
文摘Lilies are widely cultivated for cut flowers,but their large anthers carry a considerable amount of colored pollen that is dispersed easily.Studying the molecular mechanism of anther development and dehiscence could help solve this problem.LoMYB21,encoding a putative R2R3v-myb avian myeloblastosis viral oncogene homolog(MYB)transcription factor,was identified from oriental lilies(Lilium‘Siberia’).Real-time quantitative PCR analysis showed that LoMYB21 was mainly expressed in the anther,filament and stigma and had high expression during the late stages of lily anther development.LoMYB21 had transactivation activity and was located in the nucleus through yeast one-hybrid assays and transient expression in Nicotiana benthamiana.Suppression of LoMYB21 by virus-induced gene silencing(VIGS)in Lilium‘Siberia’led to anther indehiscence and reduced the expression of genes related to Jasmonate acid(JA)biosynthesis and signal transduction.Induction of LoMYB21 in DEX::LoMYB21 transgenic Arabidopsis caused procumbent inflorescences that became infertile,accompanied by higher expression of JA biosynthetic and signaling genes.These results demonstrated that JA content and signaling were abnormal in silenced lily and transgenic LoMYB21 Arabidopsis,which affected anther development.Our study indicated that LoMYB21 could regulate lily anther dehiscence through JA biosynthesis and signaling during the late stages of anther development.
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 (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 Key Research and Development Program of China(2022YFD1200500)the Fundamental Research Funds for the Central Universities(KYZZ2022004)+1 种基金the Project for Crop Germplasm Resources Conservation of Jiangsu(2021-SJ-011)the High Level Talent Project of the Top Six Talents in Jiangsu(NY-077)。
文摘Lily(Lilium spp.)is an important horticultural crop,but its use is limited due to serious pollen contamination problems.There are many studies on pollen development in model plants,but few on flower crops such as lilies.Gibberellin(GA)is a large class of hormones and plays an important role in plant vegetative growth and reproductive development.GAMYB is a group of the R2R3-MYB family upregulated by gibberellin,and plays an important role in anther development.Here,we isolated a novel GAMYB,named LoMYB65,from lily,which was closely related to the AtMYB65 and AtMYB33 in Arabidopsis.Fluorescence quantitative PCR results showed that LoMYB65 was mainly expressed in lily anthers.LoMYB65 could be activated by 288μmol·L^(-1)GA3treatment and the LoMYB65 protein was located in the nucleus and cytoplasm,and had transactivation in yeast and tobacco leaf cells.The conserved motif within 226 amino acids of the C-terminal of LoMYB65 contributed to its transactivation.Overexpression of LoMYB65 caused dwarf phenotype,unnormal tapetum development,less seeds of siliques in transgenic Arabidopsis plants,the transgenic plants showed partly male sterile.Simultaneously,silencing of LoMYB65 with VIGS(Virus Induced Gene Silencing)in lily anthers caused unnormal pollen development and reduced the pollen amount.Overexpression of LoMYB65 in Arabidopsis and silencing of LoMYB65 in lily resulted in decreased pollen counts,so we speculate that LoMYB65 may be dose-dependent.Overall,these findings suggest that LoMYB65 may play an important role in anther development and pollen formation in lily.LoMYB65 may provide a useful candidate gene for pollenless breeding of lily.
基金supported by the Science Fund for Creative Research Groups of the National Natural Science Foundation of China(32171970)the Chongqing Outstanding Scientists Project(cstc2022ycjh-bgzxm0073)the Natural Science Foundation of Chongqing,China(cstc2021jcyj-cxttX0004)。
文摘Anther dehiscence controls optimal interaction between pollen and stigma,thereby determining the successful sexual reproduction.The regulators or mechanisms of this process remain elusive.Here,two CRISPR/Cas9 mutants of a rice exocyst subunit gene SEC3A,sec3a-1 and sec3a-2,showed anther indehiscence at anthesis and male sterility at maturity.Pollen viability and germination in the mutants were partly defective,whereas their female gametes undergone a normal development.Hybrid or self-pollinated seeds could be produced by artificial pollination,suggesting potential use of a weak sec3a mutant as a female line during hybrid breeding.SEC3A is widely expressed in various tissues,including anther walls.Further results showed an excessive IAA accumulation and no endothecium lignification in sec3a-1/2 anthers.Our findings suggest that SEC3A appears to regulate anther dehiscence by modulating auxin signaling,providing insights into regulation of anther dehiscence and function of exocyst in plants.
文摘The temporal and spatial expression patterns of calmodulin mRNA in the developing tobacco anthers were investigated by in situ RNA hybridization, using digoxigening-labeled anti-RNA probe. Calmodulin mRNA was distributed in various developmental stages of tobacco anthers, but the expression level had temporal and spatial differences distinctly. During early stage of anther development, the expression level of calmodulin mRNA was significantly high, mainly distributed in epidermis, tapetum and transfusion parenchyma cells and so on. Especially, more mRNA was accumulated in the nuclei and chromosomes of microspore mother cells prior to and during meiosis. With the development of anther,mRNA was decreased gradually in the anther wall and pollen. By mature pollen stage, only a stronger positive reaction still existed in the epidermis of anther wall and transfusion parenchyma cells. The results suggest that the temporal and spatial expression of calmodulin mRNA is closely correlated with cell division,
基金supported by the China Innovative Research Team,Ministry of Education,111 Project(B14016)the National Natural Science Foundation of China(31110103915+2 种基金3117151830830014)Collaborative Innovation Center for Genetics and Development,Fudan University
文摘Rice（Oryza sativa L.） MADS3 transcription factor regulates the homeostasis of reactive oxygen species（ROS）during late anther development,and one MADS3 mutant,mads3-4,has defective anther walls,aborted microspores and complete male sterility.Here,we report the untargeted metabolomic analysis of both wild type and mads3-4 mature anthers.Mutation of MADS3 led to an unbalanced redox status and caused oxidative stress that damages lipid,protein,and DNA.To cope with oxidative stress in mads3-4 anthers,soluble sugars were mobilized and carbohydrate metabolism was shifted to amino acid and nucleic acid metabolism to provide substrates for the biosynthesis of antioxidant proteins and the repair of DNA.Mutation of MADS3 also affected other aspects of rice anther development such as secondary metabolites associated with cuticle,cell wall,and auxin metabolism.Many of the discovered metabolic changes in mads3-4 anthers were corroborated with changes of expression levels of corresponding metabolic pathway genes.Altogether,this comparative metabolomic analysis indicated that MADS3 gene affects rice anther development far beyond the ROS homeostasis regulation.
基金supported by the National Natural Science Foundation of China(No.31100225,31170289)
文摘The regulation of nutrient transportation and transformation in developing anthers is very complex. We analyzed the distribution and features of polysaccharides and lipids in the developing anthers of tomatoes using histochemical methods. Some starches appeared in the connective somatic tissue of anthers during the sporogenous cell stage. Before meiosis of the microspore mother cell,a thick polysaccharide callose wall was formed,accompanied by a reduction in the connective tissue starches. During the tetrad stage after meiosis,the polysaccharide material in the anther did not change. At the early microspore stage,the starches in the connective cells again increased,and polysaccharide material appeared in the partial intine of pollen. At the late microspore stage,a large vacuole formed that did not contain lipids or starches,and only the pollen wall contained red polysaccharides. At this stage,the connective somatic cell starch amounts decreased,and the tapetal cells changed shape and degenerated. After microspore division,abundant lipids appeared in the bicellular pollen,and starches accumulated following pollen development. As the anthers matured,many lipids and some starches accumulated in the epidermal cells. Nutrient metabolism within the tomato pollen characteristically accumulated lipids first and then starches,while the mature pollen accumulated starches and lipids simultaneously. This characteristic pattern of nutrient metabolism in tomato pollen shows species specificity among plants.
文摘Regeneration of haploid plants from wheat pollen cells by the anther culture method was long ago well accomplished by Ouyang et al. Thereafter, it has been used in wheat breeding. Hu Daofen et al. have made contribution in wheat anther cultrure breeding and a new variety obtained has been widely used in production. Successes
基金supported by the National Key R&D Program of China(2021YFD2200203)Heilongjiang Province Key R&D Program of China(GA21B010)+1 种基金Heilongjiang Touyan Innovation Team Program(Tree Genetics and Breeding Innovation Team)Heilongjiang Postdoctoral Financial Assistance(LBH-Z21097)。
文摘Doubled haploid(DH)plants have been widely used for breeding and biological research in crops.Pop ulus spp.have been used as model woody plant species for biological research.However,the induction of DH poplar plants is onerous,and limited biological or breeding work has been carried out on DH individuals or populations.In this study,we provide an effective protocol for poplar haploid induction based on an anther culture method.A total of 96 whole DH plant lines were obtained using an F1hybrid of Populus simonii×P.nigra as a donor tree.The phenotypes of the DH population showed exceptionally high variance when compared to those of half-sib progeny of the donor tree.Each DH line displayed distinct features compared to those of the other DH lines or the donor tree.Additionally,some excellent homozygous lines have the potential to be model plants in genetic and breeding studies.
基金supported by the National Natural Science Foundation of China(31771876)the Sichuan Province Science and Technology Program(2021YFYZ0011,2021YFYZ0017).
文摘Callose contributes to many biological processes of higher plants including pollen development,cell plate and vascular tissue formation,as well as regulating the transport function of plasmodesmata.The functions of callose synthase genes in maize have been little studied.We describe a maize male-sterile mutant 39(ms39)characterized by reduced plant height.In this study,we confirmed using CRISPR/Cas9 technology that a mutation in Zm00001d043909(ZmCals12),encoding a callose synthase,is responsible for the male sterility of the ms39 mutant.Compared with male-fertile plants,callose deposition around the dyads and tetrads in ms39 anthers was significantly reduced.Increased cell autophagy observed in ms39 anthers may have been due to the premature programmed cell death of tapetal cells,leading to collapse of the anther wall structure.Disordered glucose metabolism in ms39 may have intensified autophagy in anthers.Evaluation of the ms39 gene on maize heterosis by paired-crossed experiment with 11 maize inbred lines indicated that ms39 can be used for maize hybrid seed production.
基金funded by the National Key Research and Development Program of China(2021YFF1000302)Fundamental Research Funds for the Central Universities of China(06500136)the Interdisciplinary Research Project for Young Teachers of USTB(Fundamental Research Funds for the Central Universities)(FRF-IDRY-20-038).
文摘Plant cell elongation depends on well-defined gene regulations,adequate nutrients,and timely cell wall modifications.Anther size is positively correlated with the number and viability of pollen grains,while little is known about molecular mechanisms underlying anther cell elongation.Here,we found that properly activated cell elongation regulators at transcriptional levels in loss-of-function ZmMs33 mutant(ms33-6038)anthers failed to promote maize anther elongation.ZmMs33 deficiency disrupted metabolic homeostasis mainly by inhibiting both photosynthesis in anther endothecium and lipid accumulation in anther tapetum.Importantly,ms33-6038 anthers displayed ectopic,premature and excessive secondary cell wall thickening in anther middle layer,which constrained cell elongation structurally and blocked nutrient flows across different anther wall layers.The metabolic disorder was only found in ms33-6038 mutant rather than several representative male-sterility lines at transcriptional and post-translational levels.Collectively,the disordered metabolisms and blocked nutrient flows defeated the activated cell elongation regulators,and finally inhibited anther elongation and growth with a unique‘‘idling effect”in ms33-6038 mutant.
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.
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.
文摘This paper looks back to the development of plant tissue culture in China in the last century. Since 1934, tissue culture studies in China has kept up with the international development in the fields. Progress has been made by Chinese in nearly every branches of tissue culture, including in vitro organogenesis, shoot tip culture, anther culture, ovary culture, endosperm culture, protoplast culture as well as mass cell culture. On the basis of reviewing the articles written by Chinese on plant tissue culture, the internationally recognized contributions are specially mentioned. The applications of plant tissue culture to agriculture and industry in China are also introduced.
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.
