[Objective]The aim was to establish optimized system for NER isolated microspore culture of Brassica napus L.. [Method]Twenty varieties of NER of Brassica napus were grown under uncontrolled temperature and light cond...[Objective]The aim was to establish optimized system for NER isolated microspore culture of Brassica napus L.. [Method]Twenty varieties of NER of Brassica napus were grown under uncontrolled temperature and light conditions,and their isolated microspore from anthers were used as explants in vitro culture. The influencing factors of microspore culture were preliminarily studied. [Result]The difference of genotypes was important influencing factors to embryoid yield. The embryoid yield increased by supplementing with 6-BA and NAA,culturing in solid-liquid double layer medium with activated charcoal; The difference was not significant of embryoid yield between culturing in medium supplemented with colchicines and the CK. The rates of cotyledonous embryoids directly developed into normal plantlets increased through enriching with 0.1-0.2 mg/L NAA and being treated on slim illumination two days before being inducted into normal plantlets. [Conclusion]The technical system of microspore culture of restorer of new cytoplasmic male sterile （NER） was established,which and lays a foundation for accelerating genotype purification of NER introgressive line.展开更多
[ Objective] The study was to investigate the agronomic traits and breeding characteristics of genic male sterile material Mian 7AB-4- 2 in Brassica napus. [ Method] The differences in agronomic traits and polymorphis...[ Objective] The study was to investigate the agronomic traits and breeding characteristics of genic male sterile material Mian 7AB-4- 2 in Brassica napus. [ Method] The differences in agronomic traits and polymorphisms in SSR markers, between the genic male sterile material Mian 7AB-4-2 in Brassica napus and its sisterly line Mian 7AB-4-1 were investigated by hybridization and molecular identification; and the percentage of sterile individuals of Mian 7AB-4-2 and of the hybrids with its sisterly line Mian 7AB-4-1 from test cross and back cross were also studled. [ Result] Mian 7AB-4-2 was not significantly different in agronomic traits from its sisterly line Mian 7AB-4-1 at 0.05 probability level. The per- centages of sterile individuals in the pollinated fertile Mian 7AB-4-2 plants were over 60%, and that in its sisterly line Mian 7AB-4-1 was about 25%. In test crosses with other nine sterile lines, Mian 7AB-4-1 kept the percentage of sterile individuals of sterile lines over 90%, and the percentage of sterile individuals from back cross over 80%. With regard to molecular markers, Mian 7AB-4-2 and its sisterly line Mian 7AB-4-1 were different in the band number from SSR primers a2 and E10. [ Conclusion] The results indicate that Mian 7AB-4-2 is helpful for rapeseed breeding, quickening the application of new materials in field breeding.展开更多
Male sterile NEA plants were identified in progenies of the radiated 92P x Aggregate-silique in Brassica napus L. in 1993. Their progeny plants from test crossing and open pollination were 100% male sterile. The doubl...Male sterile NEA plants were identified in progenies of the radiated 92P x Aggregate-silique in Brassica napus L. in 1993. Their progeny plants from test crossing and open pollination were 100% male sterile. The double-low male sterile lines JL-4 and JL-18 were bred through successive backcrossing of the double -low variety No.4 and No. 18 in Brassica napus L.to NEA. Restorer lines 6720 and 6722 with significant heterosis in F1 were developed. The results from investigating the restoring-ntaintaining relationship and inheritance of the restorer gene indicated that JL-4 and JL-18 were different from both PolCMS and Shan 2A type, and their restorer gene was controlled by a pair of dominant genie genes.展开更多
[Objective] This study was conducted to evaluate the associations of many characters of male sterile lines in Brassica napus L.[Method]10 recessive genetic male sterile（RGMS）lines were used as parents to cross in a ...[Objective] This study was conducted to evaluate the associations of many characters of male sterile lines in Brassica napus L.[Method]10 recessive genetic male sterile（RGMS）lines were used as parents to cross in a half diallel cross method to produce 45 single cross hybrids.These 45 crosses and their 10 parents were evaluated at Guiyang during 2007-2008.[Result]The positive correlations of seed yield with plant height,pods per plant,days to flowering and days to maturity were significant.Their respective correlation coefficients（r）were 0.644,0.583,0.281 and 0.341.Highly significant and negative correlation was observed between seeds per pod and seed size（-0.533）.Direct path coefficients were observed for pods per plant（0.556 2）,seeds per pod（0.407 4）,1 000-seed weight（0.481 3）,and plant height（0.401 7）to seed yield.[Conclusion]Pods per plant,seeds per pod,1 000-seed weight and plant height were positive correlated with seed yield and showed high direct contributions to yield.Other characters including branches per plant,days to flowering,days to maturity and oil content had low contributions to seed yield.展开更多
The male sterile line is very important in the hybrid breeding program of rapeseed. This study was conducted to evaluate the combining ability of many characters of male sterile lines in Brassica napus L. Ten recessiv...The male sterile line is very important in the hybrid breeding program of rapeseed. This study was conducted to evaluate the combining ability of many characters of male sterile lines in Brassica napus L. Ten recessive genetic male sterile （RGMS） lines were used as parents to produce 45 single cross hybrids by using a half diallel cross method. These 45 crosses and their 10 parents were evaluated at Guiyang during 2007-2008. The results showed that both general combining ability （GCA） and specific combiing ability （SCA） effects were important for all characters, but additive gene effects were more predominant than non-additive gene effects. Qianyou 8AB and You 2894AB gave respective highly significant GCA effects of 230.94 and 127.65 kg-hm-2 for seed yield. Lines You 2894AB, QH303-4AB, You 157AB and You 2341AB gave highly significant GCA effects for oil content of 0.99, 1.62, 1.20 and 1.53%, respectively. The crosses among lines Qianyou 3AxQianyou 8B, Qianyou 8AxYou 2894B, You 2894AxQianyou 6B, Qianyou 8AxQH303-4B and Qianyou 8Ax Qianyou 6B gave high SCA effects of 616.29, 398.71,356.48, 394.24 and 303.79 kg hm-2 for seed yield, respectively. All these crosses also gave high seed yield indicating that these crosses could be used in the breeding program.展开更多
In Brassica napus L.(rapeseed),complete genic male sterility(GMS)plays an important role in the utilization of heterosis.Although microRNAs(miRNAs)play essential regulatory roles during bud development,knowledge of ho...In Brassica napus L.(rapeseed),complete genic male sterility(GMS)plays an important role in the utilization of heterosis.Although microRNAs(miRNAs)play essential regulatory roles during bud development,knowledge of how GMS is regulated by miRNAs in rapeseed is rather limited.In this study,we obtained a novel recessive GMS system,CN12AB.The sterile line CN12A has defects in tapetal diferentiation and degradation.llumina sequencing was employed to examine the expression of miRNAs in the buds of CN12A and the fertile line CN12B.We identifed 85 known miRNAs and 120 novel miRNAs that were expressed during rapeseed anther development.When comparing the expression levels of miRNAs between CN12A and CN12B,19 and 18 known miRNAs were found to be difrentially expressed in 0.5--1.0 mm buds and in 2.5--3.0 mm buds,respectively.Among these,the expression levels of 14 miRNAs were higher and the levels of 23 miRNAs were lower in CN12A compared with CN12B.The predicted target genes of these iferentially expressed miRNAS encode protein kinases,F-box domain-containing proteins,MADS-box family proteins,SBP-box gene family members,HD ZIP proteins,floral homeotic protein APETAL A 2(AP2)。and nuclear factor Y,subunit A.These targets have previously been reported to be involved in pollen development and male sterility,suggesting that miRNAs might act as regulators of GMS in rapeseed anthers.Furthermore,RT-qPCR data suggest that one of the dffrentially expressed miRNAs,bna-miR159,plays a role in tapetal differentiation by regulating the expression of transcription factor BnMYB101 and participates in tapetal degradation and infuences callose degradation by manipulating the expression of BnA6.These findings contibute to our understanding of the roles of miRNAs during anther development and the occurrence of GMS in rapeseed.展开更多
[Objective] This study aimed to improve the quality characters of the male sterile restorer lines of Brassica napus by recurrent selection. [Method] The dominant male sterile alleles were used as outcross medium to be...[Objective] This study aimed to improve the quality characters of the male sterile restorer lines of Brassica napus by recurrent selection. [Method] The dominant male sterile alleles were used as outcross medium to be transformed with the favourable genes from thirteen materials, and finally excellent restorer materials were bred by recurrent selection. After that, the genetic diversity, genetic gains, fatty acid chain length and unsaturarion ratios of the improved populations were analyzed. [Result] The genetic diversity and genetic gains were obviously improved by the recurrent selection. Nine materials whose oil content exceeding 50% and 14 materials whose protein content exceeding 30% were acquired, and crossed with sterile lines. The comparative experiment showed that there were 11 combinations whose oil content was obviously enhanced compared with the control. [Conclusion] Recurrent selection is an effective way to improve the quality characters of the objective pop- ulations and breed favourable male sterile restorer lines of B. napus.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
Different sterile cytoplasm types of nine cabbage cytoplasmic male sterile materials were identified by molecular marker in the study, in order to better use molecular marker to conduct the assisted breeding in the fu...Different sterile cytoplasm types of nine cabbage cytoplasmic male sterile materials were identified by molecular marker in the study, in order to better use molecular marker to conduct the assisted breeding in the future. Genomic DNA was isolated from Chinese cabbage by CTAB method. The design of two pairs of specific primers was performed on conserved flanking region of orf138 gene in the GenBank. PCR was performed with genomic DNA of the nine Chinese cabbage materials. The bands were sequenced. The homologous comparison was conducted in NCBI, and finally, the type of sterile cytoplasm was determined. The results showed that the bands were amplified only in four Chinese cabbage male sterile materials with two pairs of specific primers PUPIl and PIII/PIV, while the other five materials did not obtain the relative bands. The result was consistent with the field sterility identification. And then four molecular markers of Chinese cabbage Ogura cytoplasmic male sterility （CMS） were obtained. After conducting a homologous comparative analysis with BLAST in GenBank, it was found that the homologous degree was 100% in specific segments of tbe tbree sterility materials （L1-CI, L3-CI and L3- F1 ） and Ogu orf138 gene （GenBank accession No. ： HQ149728） of the reported broccoli Ogu CMS. The homologous degree of L1-F1 was 99% with a variation point. The type of cytoplasmic male sterility of the other five materials needed further research. Four materials of the nine were identified as the radish cytoplasmic male sterility materials and four molecular markers were obtained.展开更多
Northern blot analysis was conducted with mitochondrial RNA from seedling leaves, floral buds, and developing seeds of NCa CMS, maintainer line and fertile F1 using ten mitochondrial genes as probes. The results revea...Northern blot analysis was conducted with mitochondrial RNA from seedling leaves, floral buds, and developing seeds of NCa CMS, maintainer line and fertile F1 using ten mitochondrial genes as probes. The results revealed that 9 out of the 10 mitochondrial genes, except for atp6, showed no difference in different tissues of the corresponding materials of NCα CMS system and that they might be constitutively expressed genes. Eight genes, such as orf139, orf222, atpl, cox1, cox2, cob, rm5S, and rm26S, showed no difference among the three tissues of all the materials detected. So the expression of these eight genes was not regulated by nuclear genes and was not tissue-specific. The transcripts of atp9 were identical among different tissues, but diverse among different materials, indicating that transcription of atp9 was neither controlled by nuclear gene nor tissue-specific. Gene atp6 displayed similar transcripts with the same size among different tissues of all the materials but differed in abundance among tissues of corresponding materials and its expression might be tissue-specific under regulation of nuclear gene. Moreover, three transcripts of orf222 were detected in the floral buds of NCa cms and fertile F1, but no transcript was detected in floral buds of the maintainer line.The transcription of orf139 was similar to that of orf222 but only two transcripts of 0.8 kb and 0.6 kb were produced. The atp9 probe detected a single transcript of 0.6 kb in NCa cms and in maintainer line and an additional transcript of 1.2 kb in fertile F1. The relationship of expression of orf222, orf139, and atp9 with NCa sterility was discussed.展开更多
The unstability of cytoplasmic male sterilities (CMS) of Brassica napus Lwas stud-ied from 1989 to 1992,and the results indicated: 1. The occurrence of trace-pollen plants In sterilematerials was caused by nuclear pol...The unstability of cytoplasmic male sterilities (CMS) of Brassica napus Lwas stud-ied from 1989 to 1992,and the results indicated: 1. The occurrence of trace-pollen plants In sterilematerials was caused by nuclear polygenes of the maintainers; 2. Progenies of partial sterility typeshowed segregation of sterile, partially sterile (with trace pollen )and fertile plants; 3. High andlow temperature CMS lines were crossed and its progenies manifested complex segregation; 4.There was a tendency to increase the fertility with successive selfing of the high temperatureCMS line.展开更多
[Objectives]This study was conducted to provide a basis for the rapid identification of the drug spraying effect in early stage and the molecular mechanism of chemical hybridizing in Brassica napus L.[Methods]Quantita...[Objectives]This study was conducted to provide a basis for the rapid identification of the drug spraying effect in early stage and the molecular mechanism of chemical hybridizing in Brassica napus L.[Methods]Quantitative RT-PCR analysis showed that ALS was constitutively expressed in various tissues of 096030,including flower buds,four floral organs (calyxes,petals,stamens and pistils),roots,stems and leaves.ALS was prominently expressed in leaves and was expressed weakly in the petals and stamens.The male sterility-inducing effects of tribenuron-methyl on such two Brassica napus L.varieties as Ningyou18 and 096030 were investigated.[Results]Plants were twice sprayed with 0.2 μg/ml tribenuron-methyl on leaves.The results showed that 8-10 ml of tribenuron- methyl was applied per plant for the first time at bolting stage with 1-2 mm flower buds on 15-20 cm inflorescence,and the second spray was performed with 8-10 ml of tribenuron-methyl per plant 10 d later.The results showed that the percentage of the full sterile plants reached 100%,which lasted for the whole flowering period,and the relative seed setting rate was only about 4%.Thus,this method could fullfill the requirement of hybrid seed production in field.The in-vivo enzyme activity of acetolactate synthase (ALS) was assayed using 2 mm buds collected 3 d after spray.The results showed that 0.2 μg/ml tribenuron-methyl inhibited ALS activity.The ALS activity of Ningyou 18 (CK) and Ningyou 18 (0.2 μg/ml) was 3.20 and 1.30 μmol/(mg·h),respectively,and the ALS activity of 096030 (CK) and 096030 (0.2 μg/ml) was 3.37 and 1.25 μmol/(mg·h),respectively.The relative enzyme activity of ALS in Ningyou18 and 096030 was 40.63% and 37.23%,respectively,both of which decreased significantly.[Conclusions]These results showed that the change of ALS activity may be used as an index for quickly identifying and predicting the chemical hybridizing effect of tribenuron-methyl.展开更多
[Objective] This study was performed to screen functional genes related to the fertility conversion of thermo-sensitive genic male sterile （TGMS） lines of Brassica juncea L. [Method] A B. juncea TGMS line K121S was ...[Objective] This study was performed to screen functional genes related to the fertility conversion of thermo-sensitive genic male sterile （TGMS） lines of Brassica juncea L. [Method] A B. juncea TGMS line K121S was selected as the experimental material. The total RNAs were isolated from fertile and sterile pollens at different development stages, including mother cell stage, tetrad stage, tricellular pollen stage and maturity stage. DDRT-PCR was carried out to identify differentially expressed genes. [Result] A total of 44 differentially expressed cDNA fragments were identified with Dot blot. And seven candidate genes related to fertility conversion of K121S were screened out by BLASTN, including callose synthase gene, aldehyde dehydrogenase gene and RNA polymerase I transcription factor RRN3 gene which were differentially expressed at the transcriptional level, H＇-ATPase gene, fructose diphosphate aldolase -class I gene, teucine-rich repeat receptor-Jike serine/threonine- protein kinase gene and alkaline/neutral invertase gene, which were differentially expressed at the post-transcriptional level. [Conclusion] The results of this study will help to explain the molecular mechanism of thermo-sensitive genic male sterility of B. juncea.展开更多
Somatic hybridization is performed to obtain significant cytoplasmic male sterility （CMS） lines, whose CMS genes are derived either from the transfer of sterile genes from the mitochondrial genome of donor parent to...Somatic hybridization is performed to obtain significant cytoplasmic male sterility （CMS） lines, whose CMS genes are derived either from the transfer of sterile genes from the mitochondrial genome of donor parent to the counterpart of receptor or production of new sterile genes caused by mitochondrial genome recombination of the biparent during protoplast fusion. In this study, a novel male sterile line, SaNa-IA, was obtained from the somatic hybridization between Brassica napus and Sinapis alba. The normal anther development of the maintainer line, SaNa-IB, and the abortive process of SaNa-IA were described through phenotypic observations and microtome sections. The floral organ of the sterile line SaNa-IA was sterile with a shortened filament and deflated anther. No detectable pollen grains were found on the surface of the sterile anthers. Semi-thin sections indicated that SaNa-IA aborted in the pollen mother cell （PMC） stage when vacuolization of the tapetum and PMCs began. The tapetum radically elongated and became highly vacuolated, occupying the entire locule together with the vacuolated microspores. Therefore, SaNa-IA is different from other CMS lines, such as ogu CMS, pol CMS and nap CMS as shown by the abortive process of the anther.展开更多
基金Supported by the Key Breeding Project of Sichuan Province during National Eleventh-five Year Plan (2006YZGG25 )Key Quality Project of Sichuan Province (2006YZGG223)Foundation for Young Scientists of Sichuan Provincial Financial Breeding Project in 2008~~
文摘[Objective]The aim was to establish optimized system for NER isolated microspore culture of Brassica napus L.. [Method]Twenty varieties of NER of Brassica napus were grown under uncontrolled temperature and light conditions,and their isolated microspore from anthers were used as explants in vitro culture. The influencing factors of microspore culture were preliminarily studied. [Result]The difference of genotypes was important influencing factors to embryoid yield. The embryoid yield increased by supplementing with 6-BA and NAA,culturing in solid-liquid double layer medium with activated charcoal; The difference was not significant of embryoid yield between culturing in medium supplemented with colchicines and the CK. The rates of cotyledonous embryoids directly developed into normal plantlets increased through enriching with 0.1-0.2 mg/L NAA and being treated on slim illumination two days before being inducted into normal plantlets. [Conclusion]The technical system of microspore culture of restorer of new cytoplasmic male sterile （NER） was established,which and lays a foundation for accelerating genotype purification of NER introgressive line.
基金Supported by National High-tech R&D Program of China(2001AA241104, 2004AA241104)Key Breeding Project of Sichuan Province (200107001-1-6-1, 2006YZGG-5)Key Quality Project of Sichuan Province(2006YZGG-23)~~
文摘[ Objective] The study was to investigate the agronomic traits and breeding characteristics of genic male sterile material Mian 7AB-4- 2 in Brassica napus. [ Method] The differences in agronomic traits and polymorphisms in SSR markers, between the genic male sterile material Mian 7AB-4-2 in Brassica napus and its sisterly line Mian 7AB-4-1 were investigated by hybridization and molecular identification; and the percentage of sterile individuals of Mian 7AB-4-2 and of the hybrids with its sisterly line Mian 7AB-4-1 from test cross and back cross were also studled. [ Result] Mian 7AB-4-2 was not significantly different in agronomic traits from its sisterly line Mian 7AB-4-1 at 0.05 probability level. The per- centages of sterile individuals in the pollinated fertile Mian 7AB-4-2 plants were over 60%, and that in its sisterly line Mian 7AB-4-1 was about 25%. In test crosses with other nine sterile lines, Mian 7AB-4-1 kept the percentage of sterile individuals of sterile lines over 90%, and the percentage of sterile individuals from back cross over 80%. With regard to molecular markers, Mian 7AB-4-2 and its sisterly line Mian 7AB-4-1 were different in the band number from SSR primers a2 and E10. [ Conclusion] The results indicate that Mian 7AB-4-2 is helpful for rapeseed breeding, quickening the application of new materials in field breeding.
文摘Male sterile NEA plants were identified in progenies of the radiated 92P x Aggregate-silique in Brassica napus L. in 1993. Their progeny plants from test crossing and open pollination were 100% male sterile. The double-low male sterile lines JL-4 and JL-18 were bred through successive backcrossing of the double -low variety No.4 and No. 18 in Brassica napus L.to NEA. Restorer lines 6720 and 6722 with significant heterosis in F1 were developed. The results from investigating the restoring-ntaintaining relationship and inheritance of the restorer gene indicated that JL-4 and JL-18 were different from both PolCMS and Shan 2A type, and their restorer gene was controlled by a pair of dominant genie genes.
基金Supportd by the 11th Five Year Plan of Rapeseed Program in Guizhou Province,Scientific Contract of Guizhou N2(20053006-02-03)~~
文摘[Objective] This study was conducted to evaluate the associations of many characters of male sterile lines in Brassica napus L.[Method]10 recessive genetic male sterile（RGMS）lines were used as parents to cross in a half diallel cross method to produce 45 single cross hybrids.These 45 crosses and their 10 parents were evaluated at Guiyang during 2007-2008.[Result]The positive correlations of seed yield with plant height,pods per plant,days to flowering and days to maturity were significant.Their respective correlation coefficients（r）were 0.644,0.583,0.281 and 0.341.Highly significant and negative correlation was observed between seeds per pod and seed size（-0.533）.Direct path coefficients were observed for pods per plant（0.556 2）,seeds per pod（0.407 4）,1 000-seed weight（0.481 3）,and plant height（0.401 7）to seed yield.[Conclusion]Pods per plant,seeds per pod,1 000-seed weight and plant height were positive correlated with seed yield and showed high direct contributions to yield.Other characters including branches per plant,days to flowering,days to maturity and oil content had low contributions to seed yield.
基金Supportd by the 11th Five-year Plan of Rapeseed Program in Guizhou Province [N2 No. (2005) 3006-02-03]
文摘The male sterile line is very important in the hybrid breeding program of rapeseed. This study was conducted to evaluate the combining ability of many characters of male sterile lines in Brassica napus L. Ten recessive genetic male sterile （RGMS） lines were used as parents to produce 45 single cross hybrids by using a half diallel cross method. These 45 crosses and their 10 parents were evaluated at Guiyang during 2007-2008. The results showed that both general combining ability （GCA） and specific combiing ability （SCA） effects were important for all characters, but additive gene effects were more predominant than non-additive gene effects. Qianyou 8AB and You 2894AB gave respective highly significant GCA effects of 230.94 and 127.65 kg-hm-2 for seed yield. Lines You 2894AB, QH303-4AB, You 157AB and You 2341AB gave highly significant GCA effects for oil content of 0.99, 1.62, 1.20 and 1.53%, respectively. The crosses among lines Qianyou 3AxQianyou 8B, Qianyou 8AxYou 2894B, You 2894AxQianyou 6B, Qianyou 8AxQH303-4B and Qianyou 8Ax Qianyou 6B gave high SCA effects of 616.29, 398.71,356.48, 394.24 and 303.79 kg hm-2 for seed yield, respectively. All these crosses also gave high seed yield indicating that these crosses could be used in the breeding program.
基金This project was supported by the National Key Research Program of China(2018YFD0100500)the Major Projects for New Varieties of Genetically Modified Organisms,China(2018ZX0801109B)the Agricultural Science and Technology Innovation Project of Gansu Academy of Agricultural Sciences,China(2018GAAS04).
文摘In Brassica napus L.(rapeseed),complete genic male sterility(GMS)plays an important role in the utilization of heterosis.Although microRNAs(miRNAs)play essential regulatory roles during bud development,knowledge of how GMS is regulated by miRNAs in rapeseed is rather limited.In this study,we obtained a novel recessive GMS system,CN12AB.The sterile line CN12A has defects in tapetal diferentiation and degradation.llumina sequencing was employed to examine the expression of miRNAs in the buds of CN12A and the fertile line CN12B.We identifed 85 known miRNAs and 120 novel miRNAs that were expressed during rapeseed anther development.When comparing the expression levels of miRNAs between CN12A and CN12B,19 and 18 known miRNAs were found to be difrentially expressed in 0.5--1.0 mm buds and in 2.5--3.0 mm buds,respectively.Among these,the expression levels of 14 miRNAs were higher and the levels of 23 miRNAs were lower in CN12A compared with CN12B.The predicted target genes of these iferentially expressed miRNAS encode protein kinases,F-box domain-containing proteins,MADS-box family proteins,SBP-box gene family members,HD ZIP proteins,floral homeotic protein APETAL A 2(AP2)。and nuclear factor Y,subunit A.These targets have previously been reported to be involved in pollen development and male sterility,suggesting that miRNAs might act as regulators of GMS in rapeseed anthers.Furthermore,RT-qPCR data suggest that one of the dffrentially expressed miRNAs,bna-miR159,plays a role in tapetal differentiation by regulating the expression of transcription factor BnMYB101 and participates in tapetal degradation and infuences callose degradation by manipulating the expression of BnA6.These findings contibute to our understanding of the roles of miRNAs during anther development and the occurrence of GMS in rapeseed.
基金Supported by Science and Technology Fund of Guizhou Province[(2010)2089]Fund of Engineering Research Center of Guizhou Province[(2012)4006]Fund from Guizhou Academy of Agricultural Sciences[(2011)017]~~
文摘[Objective] This study aimed to improve the quality characters of the male sterile restorer lines of Brassica napus by recurrent selection. [Method] The dominant male sterile alleles were used as outcross medium to be transformed with the favourable genes from thirteen materials, and finally excellent restorer materials were bred by recurrent selection. After that, the genetic diversity, genetic gains, fatty acid chain length and unsaturarion ratios of the improved populations were analyzed. [Result] The genetic diversity and genetic gains were obviously improved by the recurrent selection. Nine materials whose oil content exceeding 50% and 14 materials whose protein content exceeding 30% were acquired, and crossed with sterile lines. The comparative experiment showed that there were 11 combinations whose oil content was obviously enhanced compared with the control. [Conclusion] Recurrent selection is an effective way to improve the quality characters of the objective pop- ulations and breed favourable male sterile restorer lines of B. napus.
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 Staple Vegetable Industrial Technology System(CARS-23-G37)Breeding and Industrialization Demonstration of New Varieties of Cruciferae Vegetables(Brassica pekinensis Rupr.,Raphanus sativus L.,Brassica oleracea L.)in Yunnan Province(2015BB007)Basic Research Project of Yunnan Science and Technology Plan(2017FD200)
文摘Different sterile cytoplasm types of nine cabbage cytoplasmic male sterile materials were identified by molecular marker in the study, in order to better use molecular marker to conduct the assisted breeding in the future. Genomic DNA was isolated from Chinese cabbage by CTAB method. The design of two pairs of specific primers was performed on conserved flanking region of orf138 gene in the GenBank. PCR was performed with genomic DNA of the nine Chinese cabbage materials. The bands were sequenced. The homologous comparison was conducted in NCBI, and finally, the type of sterile cytoplasm was determined. The results showed that the bands were amplified only in four Chinese cabbage male sterile materials with two pairs of specific primers PUPIl and PIII/PIV, while the other five materials did not obtain the relative bands. The result was consistent with the field sterility identification. And then four molecular markers of Chinese cabbage Ogura cytoplasmic male sterility （CMS） were obtained. After conducting a homologous comparative analysis with BLAST in GenBank, it was found that the homologous degree was 100% in specific segments of tbe tbree sterility materials （L1-CI, L3-CI and L3- F1 ） and Ogu orf138 gene （GenBank accession No. ： HQ149728） of the reported broccoli Ogu CMS. The homologous degree of L1-F1 was 99% with a variation point. The type of cytoplasmic male sterility of the other five materials needed further research. Four materials of the nine were identified as the radish cytoplasmic male sterility materials and four molecular markers were obtained.
基金This work was supported by the National High Technology R&D Project of China (No.2002AA207009) and Wuhan Dawn Project for Youth (No. 20035002016-36).
文摘Northern blot analysis was conducted with mitochondrial RNA from seedling leaves, floral buds, and developing seeds of NCa CMS, maintainer line and fertile F1 using ten mitochondrial genes as probes. The results revealed that 9 out of the 10 mitochondrial genes, except for atp6, showed no difference in different tissues of the corresponding materials of NCα CMS system and that they might be constitutively expressed genes. Eight genes, such as orf139, orf222, atpl, cox1, cox2, cob, rm5S, and rm26S, showed no difference among the three tissues of all the materials detected. So the expression of these eight genes was not regulated by nuclear genes and was not tissue-specific. The transcripts of atp9 were identical among different tissues, but diverse among different materials, indicating that transcription of atp9 was neither controlled by nuclear gene nor tissue-specific. Gene atp6 displayed similar transcripts with the same size among different tissues of all the materials but differed in abundance among tissues of corresponding materials and its expression might be tissue-specific under regulation of nuclear gene. Moreover, three transcripts of orf222 were detected in the floral buds of NCa cms and fertile F1, but no transcript was detected in floral buds of the maintainer line.The transcription of orf139 was similar to that of orf222 but only two transcripts of 0.8 kb and 0.6 kb were produced. The atp9 probe detected a single transcript of 0.6 kb in NCa cms and in maintainer line and an additional transcript of 1.2 kb in fertile F1. The relationship of expression of orf222, orf139, and atp9 with NCa sterility was discussed.
文摘The unstability of cytoplasmic male sterilities (CMS) of Brassica napus Lwas stud-ied from 1989 to 1992,and the results indicated: 1. The occurrence of trace-pollen plants In sterilematerials was caused by nuclear polygenes of the maintainers; 2. Progenies of partial sterility typeshowed segregation of sterile, partially sterile (with trace pollen )and fertile plants; 3. High andlow temperature CMS lines were crossed and its progenies manifested complex segregation; 4.There was a tendency to increase the fertility with successive selfing of the high temperatureCMS line.
基金Supported by National Natural Science Foundation of China(31571710)Earmarked Fund for China Agriculture Research System(CARS-12)
文摘[Objectives]This study was conducted to provide a basis for the rapid identification of the drug spraying effect in early stage and the molecular mechanism of chemical hybridizing in Brassica napus L.[Methods]Quantitative RT-PCR analysis showed that ALS was constitutively expressed in various tissues of 096030,including flower buds,four floral organs (calyxes,petals,stamens and pistils),roots,stems and leaves.ALS was prominently expressed in leaves and was expressed weakly in the petals and stamens.The male sterility-inducing effects of tribenuron-methyl on such two Brassica napus L.varieties as Ningyou18 and 096030 were investigated.[Results]Plants were twice sprayed with 0.2 μg/ml tribenuron-methyl on leaves.The results showed that 8-10 ml of tribenuron- methyl was applied per plant for the first time at bolting stage with 1-2 mm flower buds on 15-20 cm inflorescence,and the second spray was performed with 8-10 ml of tribenuron-methyl per plant 10 d later.The results showed that the percentage of the full sterile plants reached 100%,which lasted for the whole flowering period,and the relative seed setting rate was only about 4%.Thus,this method could fullfill the requirement of hybrid seed production in field.The in-vivo enzyme activity of acetolactate synthase (ALS) was assayed using 2 mm buds collected 3 d after spray.The results showed that 0.2 μg/ml tribenuron-methyl inhibited ALS activity.The ALS activity of Ningyou 18 (CK) and Ningyou 18 (0.2 μg/ml) was 3.20 and 1.30 μmol/(mg·h),respectively,and the ALS activity of 096030 (CK) and 096030 (0.2 μg/ml) was 3.37 and 1.25 μmol/(mg·h),respectively.The relative enzyme activity of ALS in Ningyou18 and 096030 was 40.63% and 37.23%,respectively,both of which decreased significantly.[Conclusions]These results showed that the change of ALS activity may be used as an index for quickly identifying and predicting the chemical hybridizing effect of tribenuron-methyl.
基金Supported by National Natural Science Foundation of China(31160289)Rapeseed Industry Construction Program of Department of Agriculture of Yunnan ProvinceFund for Workstation of Academician Guan Chunyun from Department of Science and Technology of Yunnan Province~~
文摘[Objective] This study was performed to screen functional genes related to the fertility conversion of thermo-sensitive genic male sterile （TGMS） lines of Brassica juncea L. [Method] A B. juncea TGMS line K121S was selected as the experimental material. The total RNAs were isolated from fertile and sterile pollens at different development stages, including mother cell stage, tetrad stage, tricellular pollen stage and maturity stage. DDRT-PCR was carried out to identify differentially expressed genes. [Result] A total of 44 differentially expressed cDNA fragments were identified with Dot blot. And seven candidate genes related to fertility conversion of K121S were screened out by BLASTN, including callose synthase gene, aldehyde dehydrogenase gene and RNA polymerase I transcription factor RRN3 gene which were differentially expressed at the transcriptional level, H＇-ATPase gene, fructose diphosphate aldolase -class I gene, teucine-rich repeat receptor-Jike serine/threonine- protein kinase gene and alkaline/neutral invertase gene, which were differentially expressed at the post-transcriptional level. [Conclusion] The results of this study will help to explain the molecular mechanism of thermo-sensitive genic male sterility of B. juncea.
基金the National Natural Science Foundation of China(31330057)the Priority Academic Program Development of Jiangsu Higher Education Institutions+1 种基金the Program of International S&T Cooperation of China(1021)the Jiangsu Province Graduate Innovation Fund(XCLX13_899),China
文摘Somatic hybridization is performed to obtain significant cytoplasmic male sterility （CMS） lines, whose CMS genes are derived either from the transfer of sterile genes from the mitochondrial genome of donor parent to the counterpart of receptor or production of new sterile genes caused by mitochondrial genome recombination of the biparent during protoplast fusion. In this study, a novel male sterile line, SaNa-IA, was obtained from the somatic hybridization between Brassica napus and Sinapis alba. The normal anther development of the maintainer line, SaNa-IB, and the abortive process of SaNa-IA were described through phenotypic observations and microtome sections. The floral organ of the sterile line SaNa-IA was sterile with a shortened filament and deflated anther. No detectable pollen grains were found on the surface of the sterile anthers. Semi-thin sections indicated that SaNa-IA aborted in the pollen mother cell （PMC） stage when vacuolization of the tapetum and PMCs began. The tapetum radically elongated and became highly vacuolated, occupying the entire locule together with the vacuolated microspores. Therefore, SaNa-IA is different from other CMS lines, such as ogu CMS, pol CMS and nap CMS as shown by the abortive process of the anther.
