Five commonly-used reference genes： ACT （actin）, UBE （ubiquitin-conjugating enzyme）, RPL2 （ribosomal protein L2）, BRP II （RNA polymerase II subunit）, and NADH （nicotinamide adenine dinucleotide） were examin...Five commonly-used reference genes： ACT （actin）, UBE （ubiquitin-conjugating enzyme）, RPL2 （ribosomal protein L2）, BRP II （RNA polymerase II subunit）, and NADH （nicotinamide adenine dinucleotide） were examined using geNorm software as reference genes for RT-qPCR. Among the tested reference genes, ACT and UBE were the most stable in all samples. In parallel, expression analysis of nitrilases in Brassica juncea var. tumida, was performed to preliminarily investigate the molecular interactions between nitrilase and clubroot development at 10, 15, 20, 25, 30, and 40 d postinoculation （dpi） with a suspension of resting spores of Plasmodiophora brassicae. The results showed that different gene expressions of nitrilases were regulated during the initial periods of clubroot development. The expression level of BjNIT1 increased sharply from 20 to 40 dpi in infected roots while there were no remarkable changes in healthy roots. From 15 to 30 dpi, the expression levels of BjNIT2 and BjNIT4 in infected roots were lower than those in non-infected roots. Finally, BjNIT2 in treatment was down approximately to control at 40 dpi. Our results suggest that BjNIT1, which promoted overproductions of auxin, might be involved in P. brassicae infection of B. juncea.展开更多
通过远缘杂交及回交将白菜的抗病基因导入到芥菜中,创制获得优质的抗根肿病长柄芥(Brassica juncea var.longepetiolata Yang et Chen)育种材料。采用人工接种鉴定和分子标记鉴定相结合法,对10份抗根肿病白菜材料进行抗性分析,筛选出1...通过远缘杂交及回交将白菜的抗病基因导入到芥菜中,创制获得优质的抗根肿病长柄芥(Brassica juncea var.longepetiolata Yang et Chen)育种材料。采用人工接种鉴定和分子标记鉴定相结合法,对10份抗根肿病白菜材料进行抗性分析,筛选出1份可作为长柄芥抗根肿病育种材料创制的抗性资源CCR21002。利用CCR21002分别作为父本、母本提供抗根肿病基因,与长柄芥进行远缘杂交,结合胚挽救技术获得正反杂交种F1,选取植株形态偏向芥菜的F1与长柄芥回交获得BC_(1),对BC_(1)植株进行形态学、细胞学分析及抗病性鉴定。结果表明,回交一代比杂种一代更难获得,胚挽救成活的13株BC_(1)中,仅筛选出1株基因型为AABB且抗根肿病的植株,经分子标记鉴定所含抗病基因为CRb,在植株形态上保留有长柄芥的大部分性状。证明经过与抗根肿病白菜远缘杂交及回交,获得对4号生理小种表现抗性的长柄芥新种质1份,可用于继续回交转育。展开更多
基金financially supported by the Natural Science Foundation of Chongqing Science and Technology Commission,China (2008BB1370)Fuling Agricultural Science Institute of Chongqing,China
文摘Five commonly-used reference genes： ACT （actin）, UBE （ubiquitin-conjugating enzyme）, RPL2 （ribosomal protein L2）, BRP II （RNA polymerase II subunit）, and NADH （nicotinamide adenine dinucleotide） were examined using geNorm software as reference genes for RT-qPCR. Among the tested reference genes, ACT and UBE were the most stable in all samples. In parallel, expression analysis of nitrilases in Brassica juncea var. tumida, was performed to preliminarily investigate the molecular interactions between nitrilase and clubroot development at 10, 15, 20, 25, 30, and 40 d postinoculation （dpi） with a suspension of resting spores of Plasmodiophora brassicae. The results showed that different gene expressions of nitrilases were regulated during the initial periods of clubroot development. The expression level of BjNIT1 increased sharply from 20 to 40 dpi in infected roots while there were no remarkable changes in healthy roots. From 15 to 30 dpi, the expression levels of BjNIT2 and BjNIT4 in infected roots were lower than those in non-infected roots. Finally, BjNIT2 in treatment was down approximately to control at 40 dpi. Our results suggest that BjNIT1, which promoted overproductions of auxin, might be involved in P. brassicae infection of B. juncea.
文摘通过远缘杂交及回交将白菜的抗病基因导入到芥菜中,创制获得优质的抗根肿病长柄芥(Brassica juncea var.longepetiolata Yang et Chen)育种材料。采用人工接种鉴定和分子标记鉴定相结合法,对10份抗根肿病白菜材料进行抗性分析,筛选出1份可作为长柄芥抗根肿病育种材料创制的抗性资源CCR21002。利用CCR21002分别作为父本、母本提供抗根肿病基因,与长柄芥进行远缘杂交,结合胚挽救技术获得正反杂交种F1,选取植株形态偏向芥菜的F1与长柄芥回交获得BC_(1),对BC_(1)植株进行形态学、细胞学分析及抗病性鉴定。结果表明,回交一代比杂种一代更难获得,胚挽救成活的13株BC_(1)中,仅筛选出1株基因型为AABB且抗根肿病的植株,经分子标记鉴定所含抗病基因为CRb,在植株形态上保留有长柄芥的大部分性状。证明经过与抗根肿病白菜远缘杂交及回交,获得对4号生理小种表现抗性的长柄芥新种质1份,可用于继续回交转育。
