【目的】探究高粱耐盐胁迫响应机制,挖掘高粱耐盐胁迫基因,为高粱耐盐育种提供理论基础。【方法】以高粱感盐品种L甜和耐盐品种石红137为供试材料,采用水培试验。待高粱植株长至三叶一心期,使用2%NaCl溶液对幼苗进行盐胁迫,分别设置0(对...【目的】探究高粱耐盐胁迫响应机制,挖掘高粱耐盐胁迫基因,为高粱耐盐育种提供理论基础。【方法】以高粱感盐品种L甜和耐盐品种石红137为供试材料,采用水培试验。待高粱植株长至三叶一心期,使用2%NaCl溶液对幼苗进行盐胁迫,分别设置0(对照)、1和24 h处理,每个处理3次重复。测定不同处理样品株高、根长、干物重、Na+含量和叶绿素相对含量(SPAD值),并依托Illumina HiSeq 2000平台进行转录组测序分析。利用FPKM方法计算基因表达量,在差异表达基因检测过程中,将差异表达倍数(fold change)≥2且FDR<0.001作为筛选标准。通过Gene Ontology和KEGG Pathway数据库对参与高粱不同时间盐胁迫差异表达基因进行分析注释。【结果】盐胁迫处理对高粱株高、根长、干物重等性状无显著影响,对钠离子含量和SPAD值影响显著。石红137株高、根长、钠离子含量和SPAD值均高于L甜。转录组测序结果鉴定得到已知基因26628个,新基因866个。石红137中的差异基因数目高于L甜。石红137中,0 h VS 1 h、0 h VS 24 h、1 h VS 24 h三组的差异基因数目分别为375、4206和3750个。感盐品种L甜中,0 h VS 1 h、0 h VS 24 h、1 h VS 24 h三组的差异基因数目分别为167、2534和1612个。GO分析共获得25个功能注释,分别为光合作用、细胞物质代谢、翻译过程以及激素合成等与盐胁迫相关的差异表达基因。KEGG分析发现盐胁迫1 h表达差异基因富集在植物激素信号转导途径,涉及脱落酸(abscisic acid,ABA)、生长素(auxin,AUX)、细胞分裂素(cytokinin,CTK)、赤霉素(gibberellins,GS)、乙烯(ethylene,ETH)过程等共71个基因。盐胁迫24 h表达差异基因富集于光合作用相关途径,涉及Lhca、Lhcb、磷酸烯醇式丙酮酸羧化酶(phosphoenolpyruvate carboxylase,PPC)、磷酸核酮糖激酶(phosphorylribonucleic kinase,PRK)等20个基因。类黄酮生物合成代谢途径差异可能是引起石红137和L甜的耐盐能力差异的原因之一,花青素还原酶(anthocyanidin reductase,ANR)和黄酮醇合成酶(flavonol synthase,FLS)参与类黄酮生物合成途径。【结论】高粱的盐胁迫过程是一个复杂的生物过程,依赖于多个基因在复杂网络中的平衡表达。盐胁迫条件下,高粱应对环境刺激受到激素信号转导和光合作用的控制。类黄酮生物合成途径在耐盐品种中起到了重要作用。展开更多
Heterosis plays an important role in the development of new crop varieties with high-yielding, good-quality and biotic/abiotic stresses while male sterile line de- velopment is the key step to determine the success of...Heterosis plays an important role in the development of new crop varieties with high-yielding, good-quality and biotic/abiotic stresses while male sterile line de- velopment is the key step to determine the success of heterosis utilization. A male sterile mutant, M207A was created in proso millet （Panicum mi/iaceurn, 2n=4x=36） for the first time using 60Co-y ray mutagenesis. Fertility identification and genetic analysis were carried out to characterize the mutant for its possible use for hetero- sis utilization in proso millet. First the sterility was investigated using both field sur- vey and indoor pollen microscopy identification. Then Pollinated by normal fertile proso millet cultivars, F1 and F2 populations from the mutant were obtained. Mean- while primary genetic analysis was also conducted using above populations in dif- ferent experimental sites, seasons and years. The results showed that the male sterile plant exhibited closed glumes, browning and dry anthers with few normal pollens. The sterility was stable and sterility rate was above 95% on average. The segregation ratio of fertile to sterile plants was 35：1 in the fertile selfing F2 popula- tion indicating that the mutant was a genic male sterility belonging to a pollen-less type controlled by a single recessive gene. The creation of the mutant, M207A can play a key role for heterosis utilization in proso millet.展开更多
文摘【目的】探究高粱耐盐胁迫响应机制,挖掘高粱耐盐胁迫基因,为高粱耐盐育种提供理论基础。【方法】以高粱感盐品种L甜和耐盐品种石红137为供试材料,采用水培试验。待高粱植株长至三叶一心期,使用2%NaCl溶液对幼苗进行盐胁迫,分别设置0(对照)、1和24 h处理,每个处理3次重复。测定不同处理样品株高、根长、干物重、Na+含量和叶绿素相对含量(SPAD值),并依托Illumina HiSeq 2000平台进行转录组测序分析。利用FPKM方法计算基因表达量,在差异表达基因检测过程中,将差异表达倍数(fold change)≥2且FDR<0.001作为筛选标准。通过Gene Ontology和KEGG Pathway数据库对参与高粱不同时间盐胁迫差异表达基因进行分析注释。【结果】盐胁迫处理对高粱株高、根长、干物重等性状无显著影响,对钠离子含量和SPAD值影响显著。石红137株高、根长、钠离子含量和SPAD值均高于L甜。转录组测序结果鉴定得到已知基因26628个,新基因866个。石红137中的差异基因数目高于L甜。石红137中,0 h VS 1 h、0 h VS 24 h、1 h VS 24 h三组的差异基因数目分别为375、4206和3750个。感盐品种L甜中,0 h VS 1 h、0 h VS 24 h、1 h VS 24 h三组的差异基因数目分别为167、2534和1612个。GO分析共获得25个功能注释,分别为光合作用、细胞物质代谢、翻译过程以及激素合成等与盐胁迫相关的差异表达基因。KEGG分析发现盐胁迫1 h表达差异基因富集在植物激素信号转导途径,涉及脱落酸(abscisic acid,ABA)、生长素(auxin,AUX)、细胞分裂素(cytokinin,CTK)、赤霉素(gibberellins,GS)、乙烯(ethylene,ETH)过程等共71个基因。盐胁迫24 h表达差异基因富集于光合作用相关途径,涉及Lhca、Lhcb、磷酸烯醇式丙酮酸羧化酶(phosphoenolpyruvate carboxylase,PPC)、磷酸核酮糖激酶(phosphorylribonucleic kinase,PRK)等20个基因。类黄酮生物合成代谢途径差异可能是引起石红137和L甜的耐盐能力差异的原因之一,花青素还原酶(anthocyanidin reductase,ANR)和黄酮醇合成酶(flavonol synthase,FLS)参与类黄酮生物合成途径。【结论】高粱的盐胁迫过程是一个复杂的生物过程,依赖于多个基因在复杂网络中的平衡表达。盐胁迫条件下,高粱应对环境刺激受到激素信号转导和光合作用的控制。类黄酮生物合成途径在耐盐品种中起到了重要作用。
基金Supported by the China Agricultural Research System(CARS-07-13.5-A3)the Special Financial Fund of Hebei(F16R03)~~
文摘Heterosis plays an important role in the development of new crop varieties with high-yielding, good-quality and biotic/abiotic stresses while male sterile line de- velopment is the key step to determine the success of heterosis utilization. A male sterile mutant, M207A was created in proso millet （Panicum mi/iaceurn, 2n=4x=36） for the first time using 60Co-y ray mutagenesis. Fertility identification and genetic analysis were carried out to characterize the mutant for its possible use for hetero- sis utilization in proso millet. First the sterility was investigated using both field sur- vey and indoor pollen microscopy identification. Then Pollinated by normal fertile proso millet cultivars, F1 and F2 populations from the mutant were obtained. Mean- while primary genetic analysis was also conducted using above populations in dif- ferent experimental sites, seasons and years. The results showed that the male sterile plant exhibited closed glumes, browning and dry anthers with few normal pollens. The sterility was stable and sterility rate was above 95% on average. The segregation ratio of fertile to sterile plants was 35：1 in the fertile selfing F2 popula- tion indicating that the mutant was a genic male sterility belonging to a pollen-less type controlled by a single recessive gene. The creation of the mutant, M207A can play a key role for heterosis utilization in proso millet.
