烟草HPL基因的生物信息学分析与抗逆功能分析

Stress Resistance Function and Bioinformatics Analysis of HPL Gene in Nicotiana tabacum

为了探究烟草脂氢过氧化物裂解酶(HPL)的功能,以烟草叶片cDNA为模板克隆HPL基因,获得过表达NtHPL转基因烟株进行抗逆性功能研究。结果表明,NtHPL基因包含1个内含子和2个外显子,其cDNA全长为1494 bp;NtHPL蛋白的C-端具有细胞色素P450蛋白家族的保守序列,N-端含有叶绿体转运肽;NtHPL在根、茎、叶中均有表达,叶中的表达量最高;机械损伤、茉莉酸甲酯(MeJA)、水杨酸(SA)、乙烯、脱落酸(ABA)和干旱胁迫能不同程度地提高叶片NtHPL的表达水平;干旱处理后,NtHPL转基因烟株的脯氨酸含量明显高于野生型K326,丙二醛(MDA)含量、过氧化物酶(POD)和超氧化物歧化酶(SOD)活性低于野生型K326。以上结果表明,NtHPL能够提高烟株的抗旱能力,可用于烟草抗旱品种的培育。

In order to explore the function of lipid hydroperoxidelyase(HPL)in tobacco,NtHPL gene was cloned from leaves,and the transgenic tobacco plants with overexpression of NtHPL were obtained to research the stress resistance function of NtHPL.NtHPL gene contained an intron and two exons,and the full-length of its cDNA was 1494 bp.The C-terminal of NtHPL protein included the conservative sequence of the cytochrome P450 protein family,and N-terminal contained a chloroplast transit peptide.The expression of NtHPL could be found in tobacco roots,stems and leaves,and its expression was the highest in leaves.Mechanical damage,methyl jasmonate(MeJA),salicylic acid(SA),ethylene,abscisic acid(ABA)and drought stress could increase the expression level of NtHPL in leaves.Under drought stress,proline content was significantly higher in the transgenic tobacco plants with overexpression of NtHPL than that in the wild type K326,and malondialdehyde(MDA)content,peroxidase(POD)and superoxide dismutase(SOD)activities were lower than those in the wild type K326.These results showed that the NtHPL could improve the drought-resistant ability of tobacco plants,and it could be used for the breeding of tobacco drought-resistant varieties.