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含异位表达花生AhNCED1基因的拟南芥提高耐渗透胁迫能力
  • 期刊名称:作物学报
  • 时间:0
  • 页码:1440-1449
  • 语言:中文
  • 分类:S565.2[农业科学—作物学]
  • 作者机构:[1]仲恺农业工程学院生命科学学院,广东广州510225, [2]仲恺农业工程学院教学科研基地,广东广州510225
  • 相关基金:本研究由国家自然科学基金项目(30800077)资助
  • 相关项目:抗旱性不同花生品种中AhNCED1基因启动子的克隆及活性分析
中文摘要:

AhNCED1是干旱胁迫下调控花生ABA生物合成的关键基因。以pCAMBIA1301为基本双元表达载体,分别构建CaMV35S启动子和拟南芥AtNCED3基因启动子(AtNCED3p)驱动花生AhNCED1基因的2个植物双元表达载体p35S::ORF和pAtNCED3p::ORF,通过根癌农杆菌介导法将上述两个表达载体分别转化野生型和129B08/nced3突变体拟南芥,经潮霉素筛选和PCR鉴定分别获得35S::ORF-WT和A3p::ORF-B08转基因植株,RT-PCR证实花生AhNCED1基因已在转基因植株中稳定表达,并对野生型、129B08/nced3突变体和转基因拟南芥进行外源ABA敏感性和耐渗透胁迫能力分析。结果表明,129B08/nced3突变体对外源ABA的敏感性下降,而花生AhNCED1基因在拟南芥中的异位表达提高了对外源ABA的敏感性。在山梨醇胁迫下,129B08/nced3突变体种子的相对萌发率明显低于野生型的,而A3p::ORF-B08转基因拟南芥种子的相对萌发率与野生型的相当,显著高于129B08/nced3突变体的,且300mmolL–1山梨醇胁迫下,35S::ORF-WT转基因拟南芥种子的相对萌发率明显高于野生型的。在300mmolL–1山梨醇胁迫下,129B08/nced3突变体幼苗叶片高度黄化,根的形成和幼苗生长受到严重抑制,而A3p::ORF-B08转基因突变体与野生型相似,叶片仅轻度黄化,幼苗生长势良好;35S::ORF-WT转基因植株幼苗生长未受明显影响。这些结果说明,拟南芥129B08/nced3突变体对山梨醇诱导的非离子渗透胁迫有超敏性,异位表达花生AhNCED1基因能恢复该突变体对山梨醇的超敏性,提高拟南芥的耐渗透胁迫能力。

英文摘要:

The phytohormone abscisic acid (ABA) is a key regulator of seed development, root growth, stomatal aperture in higher plants and it is also involved in adaptation of plants to various stresses. The oxidative cleavage of cis-epoxycarotenoids catalyzed by nine-cis-epoxycarotenoid dioxygenase (NCED) is considered to be the rate-limiting step in ABA biosynthesis in higher plants. The AhNCED1 gene plays a vital role in the regulation of ABA biosynthesis in peanut plants in response to drought stress. Two binary vectors, p35S::ORF and pAtNCED3p::ORF, were established which harbored the AhNCED1 gene, respectively, driven by the CaMV 35S promoter originated from pCAMBIA1301 and the AtNCED3 gene promoter from wild type Arabidopsis. Wild type and 129B08/nced3 mutant Arabidopsis plants were separately transformed with Agrobacterium harboring p35S::ORF or pAtNCED3p::ORF vectors, generating 35S::ORF-WT and A3p::ORF-B08 transgenic plants, respectively, after hygromycin screening and PCR detection. The stable expression of AhNCED1 gene in Arabidopsis plants was confirmed by duplex RT-PCR performance. Wild type, 129B08/nced3 mutant and transgenic Arabidopsis plants were subsequently tested for sensitivity to exogenous ABA and tolerance to osmotic stress. The results showed that the ABA sensitivity of 129B08/nced3 mutant declined, and that of Arabidopsis plants ectopically expressing the AhNCED1 gene increased. Under sorbitol stress, the relative germination rate of 129B08/nced3 mutant seeds was far lower than that of wild type seeds; however, the relative germination rate of A3p::ORF-B08 transgenic seeds was close to that of wild type seeds, significantly higher than that of 129B08/nced3 mutant seeds. The relative germination rate of 35S::ORF-WT transgenic seeds was higher than that of wild type seeds at the treatment of 300 mmol L–1 sorbitol. Under 300 mmol L–1 sorbitol stress, leaf of 129B08/nced3 mutant plants was highly chlorotic, and root formation and seedling growth were severel

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