耐盐水稻长毛谷的生理测定及其全基因组测序分析

Physiological determination and whole genome sequencing analysis of a salt-tolerant rice Changmaogu

  • 摘要: 【目的】探讨耐盐水稻长毛谷的苗期耐盐性,并对其全基因组测序数据进行分析,以期阐明长毛谷的耐盐生理机制,为挖掘优质耐盐水稻品种及改良盐碱地提供参考。【方法】以Pokkali(耐盐对照)、IR29(盐敏感对照)及前期筛选出的耐盐水稻长毛谷为试验材料,设5种盐浓度(0、80、100、120和150 mmol/L)处理,测定3份水稻材料苗期的9项表型指标和13项生理指标,并基于全基因组测序数据探讨长毛谷的耐盐生理机制。【结果】盐胁迫下长毛谷的各项表型指标受盐浓度变化的影响最小,抗氧化酶活性更强,其他活性物质、渗透调节物质含量更高,盐胁迫对其造成的细胞膜损伤更小,对盐胁迫具有较好的耐受性。相关分析结果显示,长毛谷幼苗的生物量积累与苗高、根长、根尖数、根表面积和根体积呈极显著正相关(P<0.01,下同),与丙二醛(MDA)和过氧化氢(H2O2)含量呈极显著负相关,且超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)活性及可溶性蛋白和谷胱甘肽(GSH)含量6个生理指标间两两呈极显著正相关。通过将30个已知水稻耐盐基因蛋白序列与长毛谷蛋白序列进行比对,发现长毛谷中有28个耐盐基因的相似度和同源性达到或高于98%,而OsP5CSOsBADH1基因的相似度分别为96%和90%;对于OsP5CS基因,长毛谷比参考蛋白序列多30个氨基酸;对于OsBADH1基因,长毛谷比参考蛋白序列少17个氨基酸,同时存在4个位点的氨基酸差异,推测该差异可能与耐盐性相关。【结论】盐胁迫下长毛谷的耐盐表现优异,且2个耐盐基因的蛋白序列与参考基因存在差异。

     

    Abstract: 【Objective】The purpose of the study was to investigate the salt tolerance of salt-tolerant rice Changmaogu at seedling stage and to analyze its whole genome sequencing data,in order to clarify the salt-tolerant physiological mechanism of Changmaogu and to provide reference for the excavation of high-quality salt-tolerant rice varieties and the improvement of saline-alkali land.【Method】Pokkali (salt-tolerant control),IR29 (salt-sensitive control) and the prescreened salt-tolerant rice Changmaogu were selected as the research objects.Five salt concentrations (0,80,100,120and 150 mmol/L) were set as treatments.At seedling stage,9 phenotypic indexes and 13 physiological indexes of three rice materials were observed,and the salt-tolerant physiological mechanism of Changmaogu was investigated based on whole genome sequencing data.【Result】Under salt stress,the phenotypic indexes of Changmaogu were least affected by the change of salt concentration.The activity of antioxidant enzymes was stronger,the content of other active substances and osmotic regulatory substances was higher,the damage of cell membrane caused by salt stress was smaller,and it had relatively good tolerance to salt stress.Correlation analysis results showed that the biomass accumulation of Changmaogu seedlings was extremely significantly positively correlated with seedling height,root length,number of root tips,root surface area and root volume (P<0.01,the same below),and extremely significantly negatively correlated with the con-tents of malonaldehyde(MDA) and hydrogen peroxide(H2O2).And the six physiological indexes of the activities of superoxide dismutase(SOD),peroxidase(POD),catalase(CAT),ascorbate peroxidase(APX) and the contents of soluble pro-tein and glutathione(GSH) were extremely significantly positively correlated with each other.By comparing the protein sequences of 30 known salt-tolerant genes in rice with those of Changmaogu,it was found that the similarity and homology of 28 salt-tolerance genes in Changmaogu reached 98%or above,while the similarity of OsP5CS and OsBADH1genes were 96%and 90%respectively.For OsP5CS gene,Changmaogu contained 30 amino acids more than the reference protein sequence.For OsBADH1 gene,Changmaogu was 17 amino acids less than the reference protein sequence,and there were four amino acid sites of difference at the same time,suggesting that the differences might be related to salt tolerance.【Conclusion】Changmaogu has excellent salt tolerance under salt stress,and the protein sequences of two salttolerant genes are different from those of the reference genes.

     

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