To determine differential expression of genie male sterility A/B lines in Chinese cabbage-pak-choi (Brassica campestris ssp. chinensis Makino var. communis Tsen et Lee), we used the RNA fingerprinting technique, cDNA-...To determine differential expression of genie male sterility A/B lines in Chinese cabbage-pak-choi (Brassica campestris ssp. chinensis Makino var. communis Tsen et Lee), we used the RNA fingerprinting technique, cDNA-AFLP analysis, in different developmental stages and different tissues. While no obvious differential expressions were observed in rosette leaves, florescence leaves, and scapes, some differential expressions were found in alabstrums of A/B lines and among leaves, scapes and alabstrums. We analyzed the al-abstrums collected in different developmental stages with 10 primer combinations. We got a unique band between middle size alabstrums and large alabstrums in B line in one of the ten pair primers, and in another one pair, one band reflecting a higher gene-expression level in A line than that in B line was obtained. No unique bands were found with the other primer combinations. The bands reflecting different gene-expression level were confirmed by Northern hybridization. The results indicated that cDNA-AFLP was a suitable tool for studying differential expression of genie male sterility in plants. SDS-polyacrylamide gel electrophoresis patterns of soluble proteins further verified the difference in A/B lines.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
Seedlings of wheat (Triticum aestivum L.), Chinese cabbage (Brassica campestris L.) and mung bean (Phaseolus radiatus L.) were grown for 30 days in sterile sand media with 6 N treatments, i.e. NH4+-N, glycine-N, 3 dif...Seedlings of wheat (Triticum aestivum L.), Chinese cabbage (Brassica campestris L.) and mung bean (Phaseolus radiatus L.) were grown for 30 days in sterile sand media with 6 N treatments, i.e. NH4+-N, glycine-N, 3 different ratios of glycine-N:NH4+-N (NH4+-N was labeled with 15N) and a control receiving no N, to assess the importance of amino acids in excessive N nutrition along with inorganic N interactions. The contribution of nitrogen derived from glycine-N to total plant N was investigated. The total plant N of the three species treated with N was significantly greater (P < 0.05) than the control treatment. Also, seedlings from all the three species had significantly more total N as NH4+-N (P < 0.05) than at least two of the four treatments with glycine-N. However, for all species, differences in total N among treatments with a mixture of glycine-N and NH4+-N were mostly not significant. The contribution of N derived from glycine-N to plant total N content for all species increased with increasing glycine-N:NH4+-N ratio in the treatment solution. These results indicated that agricultural plants could effectively use organic nitrogen from organic nitrogen sources (e.g., glycine) and from organic and inorganic N mixtures (e.g., a glycine-N and NH4+-N mix). There were also genotypic differences in glycine-N and NH4+-N uptake by agricultural species.展开更多
基金supported by the National Natural Science Foundation of China(39670512)
文摘To determine differential expression of genie male sterility A/B lines in Chinese cabbage-pak-choi (Brassica campestris ssp. chinensis Makino var. communis Tsen et Lee), we used the RNA fingerprinting technique, cDNA-AFLP analysis, in different developmental stages and different tissues. While no obvious differential expressions were observed in rosette leaves, florescence leaves, and scapes, some differential expressions were found in alabstrums of A/B lines and among leaves, scapes and alabstrums. We analyzed the al-abstrums collected in different developmental stages with 10 primer combinations. We got a unique band between middle size alabstrums and large alabstrums in B line in one of the ten pair primers, and in another one pair, one band reflecting a higher gene-expression level in A line than that in B line was obtained. No unique bands were found with the other primer combinations. The bands reflecting different gene-expression level were confirmed by Northern hybridization. The results indicated that cDNA-AFLP was a suitable tool for studying differential expression of genie male sterility in plants. SDS-polyacrylamide gel electrophoresis patterns of soluble proteins further verified the difference in A/B lines.
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.
文摘通过制作石蜡切片研究了菜薹[Brassi cacampestris L.ssp.chinensis （L.） Makinovar.utilis Tsen et Lee]早熟品种‘油青四九’和晚熟品种‘油青甜菜心80天’的花芽分化过程，结果表明，当展开2-3片真叶时花芽分化开始启动。用已报道的拟南芥Flowering locus C（FLC）基因和FRIGIDA（FRI）基因的保守区域设计引物，通过RT-PCR的方法从两个菜薹品种中均克隆得到了两个决定开花的关键基因，并命名为BrcuFLC（GenBank登录号为EF138603）和BrcuFRI（GenBank登录号为EU700362）。半定量式RT-PCR表达分析表明，BrcuFLC基因在早、晚熟菜薹品种的不同发育时期表达存在差异，表达量随真叶数增加而逐步减少，但在晚熟品种中BrcuFLC表达量降低幅度小；BrcuFRI则在早、晚熟品种的所有阶段表达都较低。BrcuFLC在菜薹不同部位表达的情况不同，在茎、叶中的表达强，花次之，根中表达较弱；而Brcu-FRI在早、晚熟品种根中的表达量明显高于其它3个部位。
基金1Project supported by the National Natural Science Foundation of China (No. 30370838) and the Specialized ResearchFund for the Doctoral Program of Higher Education (No. 20030335076).
文摘Seedlings of wheat (Triticum aestivum L.), Chinese cabbage (Brassica campestris L.) and mung bean (Phaseolus radiatus L.) were grown for 30 days in sterile sand media with 6 N treatments, i.e. NH4+-N, glycine-N, 3 different ratios of glycine-N:NH4+-N (NH4+-N was labeled with 15N) and a control receiving no N, to assess the importance of amino acids in excessive N nutrition along with inorganic N interactions. The contribution of nitrogen derived from glycine-N to total plant N was investigated. The total plant N of the three species treated with N was significantly greater (P < 0.05) than the control treatment. Also, seedlings from all the three species had significantly more total N as NH4+-N (P < 0.05) than at least two of the four treatments with glycine-N. However, for all species, differences in total N among treatments with a mixture of glycine-N and NH4+-N were mostly not significant. The contribution of N derived from glycine-N to plant total N content for all species increased with increasing glycine-N:NH4+-N ratio in the treatment solution. These results indicated that agricultural plants could effectively use organic nitrogen from organic nitrogen sources (e.g., glycine) and from organic and inorganic N mixtures (e.g., a glycine-N and NH4+-N mix). There were also genotypic differences in glycine-N and NH4+-N uptake by agricultural species.
