Sesame （Sesamue indicum L.） is one of the most important oilseed crops with high oil yield. Here, we described a simple and efficient method for constructing a normalized cDNA library from a high oil content cultiva...Sesame （Sesamue indicum L.） is one of the most important oilseed crops with high oil yield. Here, we described a simple and efficient method for constructing a normalized cDNA library from a high oil content cultivar of sesame Zhongzhi 14, during its oil accumulation stages. It combined switching mechanism at 5？end of RNA transcript （SMART） technique and duplex-specific nuclease （DSN） normalization methods. Double-stranded cDNAs were synthesized from mRNAs, processed by normalization and Sfi I restriction endonuclease, and finally the cDNAs were ligated to pDNR-LIB vector. The ligation mixture was transformed into Escherichia coli DH10B by electroporation. The capacity of the library was 1.0？06 clones in this library. Gel electrophoresis results indicated the fragments ranged from 700 to 2 000 bp, with the average size of 1 800 bp. Random picking clones showed that the recombination rate was 100%. The results showed that the cDNA library constructed successfully was a full-length library with high quality, and could be used to screen the genes related to development of oil synthesis.展开更多
Cross combinations from six rapeseed cultivars and lines were evaluated under waterlogging stress condition in order to understand the genetic mechanism of waterlogging tolerance of Brassica napus L.and provide reason...Cross combinations from six rapeseed cultivars and lines were evaluated under waterlogging stress condition in order to understand the genetic mechanism of waterlogging tolerance of Brassica napus L.and provide reasonable improvement programs.There were six germination traits investigated on combining ability and heritability using complete diallel crossing method designed for 30 combinations from those six cultivars and lines.The traits included relative root length,stem length,fresh weight per plant,survival rate,electrical conductivity,and vigor index.After flooding treatment,the six traits of parents and F1 were analyzed.The general combining ability（GCA） and special combining ability（SCA） of germination traits were analyzed using Griffing I method.Among 30 cross combinations,the GCA was significantly different among six waterlogging resistance traits.The SCA of these traits was significantly different except the SCA of electrical conductivity.As a representative trait of waterlogging tolerance in rapeseed,relative vigor index had the highest narrow heritability and relatively low broad heritability.The cultivars Zhongshuang 9 and P79 had higher tolerance potential to waterlogging stress.It can be concluded that combining ability and genetic effects of relative vigor index during germination stage could be used to identify the waterlogging tolerance of rapeseed in breeding program.展开更多
基金supported by the National Basic Research Program of China (2011cb109305)the Genetically Modified Organisms Breeding Major Projects, China (2009zx08004-002B)+1 种基金the Open Project Program of Key Laboratory for Oil Crops Biology, the Ministry of Agriculture, China (200703)the Foundation of Oil Crops Research Institute, Chinese Academy of Agricultural Sciences
文摘Sesame （Sesamue indicum L.） is one of the most important oilseed crops with high oil yield. Here, we described a simple and efficient method for constructing a normalized cDNA library from a high oil content cultivar of sesame Zhongzhi 14, during its oil accumulation stages. It combined switching mechanism at 5？end of RNA transcript （SMART） technique and duplex-specific nuclease （DSN） normalization methods. Double-stranded cDNAs were synthesized from mRNAs, processed by normalization and Sfi I restriction endonuclease, and finally the cDNAs were ligated to pDNR-LIB vector. The ligation mixture was transformed into Escherichia coli DH10B by electroporation. The capacity of the library was 1.0？06 clones in this library. Gel electrophoresis results indicated the fragments ranged from 700 to 2 000 bp, with the average size of 1 800 bp. Random picking clones showed that the recombination rate was 100%. The results showed that the cDNA library constructed successfully was a full-length library with high quality, and could be used to screen the genes related to development of oil synthesis.
基金supported by the National High-Tech Research and Development Program of China (863 Program,2006AA10Z1C2)the Key Technologies R&D Program of China during the 10th Five-Year Plan period (2009BADA8B01,2110BAD01B09)the Natural Science Foundation of Hubei Province,China(2009CDA089)
文摘Cross combinations from six rapeseed cultivars and lines were evaluated under waterlogging stress condition in order to understand the genetic mechanism of waterlogging tolerance of Brassica napus L.and provide reasonable improvement programs.There were six germination traits investigated on combining ability and heritability using complete diallel crossing method designed for 30 combinations from those six cultivars and lines.The traits included relative root length,stem length,fresh weight per plant,survival rate,electrical conductivity,and vigor index.After flooding treatment,the six traits of parents and F1 were analyzed.The general combining ability（GCA） and special combining ability（SCA） of germination traits were analyzed using Griffing I method.Among 30 cross combinations,the GCA was significantly different among six waterlogging resistance traits.The SCA of these traits was significantly different except the SCA of electrical conductivity.As a representative trait of waterlogging tolerance in rapeseed,relative vigor index had the highest narrow heritability and relatively low broad heritability.The cultivars Zhongshuang 9 and P79 had higher tolerance potential to waterlogging stress.It can be concluded that combining ability and genetic effects of relative vigor index during germination stage could be used to identify the waterlogging tolerance of rapeseed in breeding program.
