优质牧草扁蓿豆SKIP同源基因的克隆和功能鉴定

发布时间：2018-07-03 05:59

  本文选题：扁蓿豆 + MrSKIP基因　； 参考：《内蒙古大学》2017年硕士论文

【摘要】：本研究用含20%PEG 6000的MS液体培养基模拟生理脱水胁迫处理扁蓿豆幼苗,以上述胁迫处理6 h的扁蓿豆幼苗叶片总RNA为模板进行反转录,运用RACE技术和RT-PCR方法,克隆得到扁蓿豆抗逆相关RNA剪切因子MrSKIP基因cDNA全长。序列分析结果显示,MrSKIP基因cDNA全长为2280bp,含有一个1833bp的完整开放阅读框,序列推测编码610个氨基酸含一条S-N-W-K-N的多肽,分子量为68.86145 KDa,等电点为8.95,生物信息学分析表明MrSKIP含有该家族的典型结构域SNW_SKIP。进一步克隆得到了扁蓿豆基因组的MrSKIP基因,序列分析表明,该基因无内含子。利用tasiRNA技术和发根农杆菌转化法,在截形苜蓿108R根部干扰MrSKIP同源基因的表达,对转化外源基因的截型苜蓿根系进行干旱、高盐和低温胁迫处理,鉴定MrSKIP基因的功能,结果显示胁迫处理下干扰MrSKIP同源基因的的转基因根系样品,其根长和根鲜重均显著小于对照。研究结果初步表明MrSKIP基因表达量的降低影响植物对干旱、低温和高盐胁迫的耐受性,特别对干旱胁迫耐受性的降低尤为显著。
[Abstract]:In this study, a MS liquid medium containing 20 and PEG 6000 was used to simulate physiological dehydration stress to treat almond bean seedlings. The leaf total RNA of almond bean seedlings treated with above stress for 6 h was used as template for reverse transcription. Race technique and RT-PCR method were used. The full-length cDNA of rskIP gene was obtained by cloning. Sequence analysis showed that the full length cDNA of Mr skp gene was 2280bp. it contained a complete open reading frame of 1833bp. The sequence was deduced to encode 610 amino acids containing a polypeptide of S-N-W-K-N. The molecular weight is 68.86145 KDaand the isoelectric point is 8.95. The bioinformatics analysis shows that MrskIP contains the typical domain SNWSKIPs of this family. The MRSKIP gene was further cloned and sequenced, which showed that the gene had no intron. Using tasiRNA technique and Agrobacterium rhizogenes transformation method, the expression of MRSKIP homologous gene was interfered at the root of truncated alfalfa (108R). The root system of truncated alfalfa transformed with exogenous gene was treated with drought, high salt and low temperature stress to identify the function of MRSKIP gene. The results showed that the root length and fresh weight of the transgenic root samples which interfered with the MRSKIP homologous gene under stress were significantly lower than that of the control. The results showed that the decreased expression of MrskIP gene affected the tolerance of plants to drought, low temperature and high salt stress, especially to drought stress.
【学位授予单位】：内蒙古大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：Q943.2

【相似文献】

相关期刊论文 前10条

1 王照兰;杨持;杜建材;胡卉芳;赵丽丽;毛小涛;;不同生态型扁蓿豆光合特性和光适应能力[J];生态学杂志;2009年06期

2 李志勇;师文贵;李鸿雁;李兴酉;刘磊;;内蒙古3种生态型扁蓿豆遗传多样性与亲缘关系的分析[J];西北植物学报;2009年12期

3 李小安;;低温吸胀对不同扁蓿豆种子发芽特性的影响[J];青海大学学报(自然科学版);2011年02期

4 李小安;;低温胁迫对不同扁蓿豆种子内游离脯氨酸和可溶性糖含量的影响[J];青海科技;2011年02期

5 张宇;石凤翎;云娜;高霞;伊风艳;蔡丽艳;;扁蓿豆冷诱导基因差异表达分析[J];西北植物学报;2013年01期

6 师文贵;李志勇;李鸿雁;赵成贵;李兴酉;刘磊;;扁蓿豆不同分布区野生居群形态多样性研究[J];西北植物学报;2009年05期

7 李小安;;低温胁迫对青藏扁蓿豆与和田苜蓿种子游离脯氨酸和可溶性糖含量的影响[J];青海大学学报(自然科学版);2011年04期

8 黄美娟，黄绍兴，朱o，

本文编号：2092657