营养液漂浮育苗棉苗根系适应水生环境的机理研究

doi:10.3864/j.issn.0578-1752.2014.17.005

摘要/Abstract

摘要： 【目的】棉花营养液漂浮育苗是一种新型的育苗技术，但棉花根系对水生环境的适应机理尚不清楚。作者从根系的结构、蛋白质组学以及关键基因表达等方面研究棉花根系的适应机理。【方法】以基质育苗为对照，用透射电镜和光学显微镜观察营养液漂浮育苗不同苗期棉苗主根尖的超微与显微结构，以蛋白质双向电泳技术分析营养液漂浮育苗3叶1心期棉苗水生根的蛋白质差异，并用实时荧光定量PCR技术进行营养液漂浮育苗不同苗期棉苗水生根以及3叶1心期棉苗不同部位关键基因即乙醇脱氢酶基因与烯醇酶基因的表达验证。【结果】营养液漂浮育苗棉苗主根中各个时期均出现了通气组织。漂浮育苗的皮层薄壁细胞体积较小。漂浮育苗棉苗主根的韧皮部生长基本正常，而木质部生长出现弱化，其导管直径明显比对照小，导管占中柱的比例也比对照小。1叶1心期，漂浮育苗棉苗主根尖细胞中出现酚类物质。2叶1心期，漂浮育苗的主根尖皮层细胞之间存在明显的细胞间隙。3叶1心期，漂浮育苗的主根尖中出现淀粉粒。4叶1心期，漂浮育苗棉苗的主根尖中出现含晶细胞，同时，细胞中出现明显的质壁分离现象。分析漂浮育苗条件下3叶1心期棉苗的水生根与对照根系蛋白质双向电泳的结果，总计28个差异蛋白被鉴定出来，28个差异蛋白包括20个非冗余蛋白。对这些蛋白进行功能分类，主要涉及新陈代谢、细胞结构、细胞防卫、能量代谢、蛋白质合成、细胞生长等，涉及的主要代谢途径为：糖酵解、乙醇发酵、三羧酸循环等途径。营养液漂浮育苗棉苗水生根中的乙醇脱氢酶2a基因从2叶1心期至5叶1心期与对照相比均有显著差异。水生根中1叶1心期至4叶1心期相对表达量呈上升趋势，4叶1心期至5叶1心期呈急剧下降趋势，在3叶1心期漂浮育苗棉苗水生根中的乙醇脱氢酶2a基因相对表达量与漂浮棉苗茎、叶相比有显著差异。营养液漂浮育苗棉苗水生根中的烯醇酶基因从1叶1心期至4叶1心期的相对表达量比对照均有显著差异，其相对表达量从1叶1心期至5叶1心期呈逐渐下降的趋势，在3叶1心期漂浮育苗棉苗水生根中的烯醇酶基因相对表达量与漂浮棉苗茎、叶相比有显著差异。【结论】在营养液漂浮育苗环境下，棉苗水生根通过根系结构及基因表达的变化来适应水生环境，是棉花对水生环境适应机理。

关键词: 棉花 , 营养液漂浮育苗 , 根系结构 , 蛋白质组 , 基因表达分析 , 适应机理

Abstract: 【Objective】Floating nursing seeding in nutrient water-bed (FNSNW) is a newly developed nursing seedling technique of cotton, but the adaptation mechanism to aquatic environment of roots is not clear. In this study, the possible mechanism of cotton roots adapting to FNSNW was analyzed using root structure, proteomics and the key gene expression.【Method】The matrix seedling method was used as control, ultrastructure and microstructure of main root tip of cotton seedling bred with FNSNW at different seedling stages were observed by transmission electron microscope and optical microscope. Protein difference between the control and aquatic root at the 3-leaf and 1-heart stage in FNSNW was analyzed using two-dimensional electrophoresis analysis. The expression of key genes including alcohol dehydrogenase gene and enolase gene for cotton seedling aquatic roots at different seedling stages and for different tissues of the same seeding at the 3-leaf and 1-heart stage in FNSNW was confirmed by real-time fluorescence quantitative PCR technology.【Result】In floating nursing seeding in nutrient water-bed (FNSNW), main roots of cotton seedling were found with appearance of aerenchyma. Compared with control, the cortical parenchyma cells of main root in FNSNW were smaller, and the growth of the xylem was weaker. Especially, the conduit diameter and the ratio of the conduit in stele were significantly smaller than the control. For tip cells of main root in FNSNW, there was phenol material at the 1-leaf and 1-heart stage, cortical cell gap at the 2-leaf and 1-heart stage, starch grain at the 3-leaf and 1-heart stage, and crystal cells with plasmolysis in the 4 leaf and 1-heart stage. As the protein two-dimensional electrophoresis results of aquatic roots and control roots at the 3-leaf and 1-heart stage, there were 28 differentially expressed proteins including 20 non-redundant proteins. Protein functional classification indicated that these proteins were related with metabolism, cell structure, cell defense, energy metabolism, protein synthesis and cell growth, and mainly involved in metabolic pathway including glycolysis, ethanol fermentation, and tricarboxylic acid cycle. Compared with control, the expression of alcohol dehydrogenase 2a gene of aquatic root in FNSNW was significantly different with first increase from the 2-leaf and 1-heart stage to the 4-leaf and 1-heart stage and then decrease from the 4-leaf and 1-heart stage to the 5-leaf and 1-heart stage, and the expression of enolase gene was significantly different with decrease from the 1-leaf and 1-heart stage to the 5-leaf and 1-heart stage. For the same cotton seedling in FNSNW at the 3-leaf and 1-heart stage, the relative expressions of these two genes of aquatic roots was significantly different from stem and leaf.【Conclusion】In FNSNW environment, seedlings adapt to the aquatic environment through changes in root structure and gene expression, which may be the adaption mechanism for cotton seedlings bred by FNSNW.

Key words: cotton , floating nursing seedling in nutrient water-bed (FNSNW) , roots structure , proteomics , gene expression analysis , adaptation mechanism

张昊，陈金湘，刘海荷，周仲华，王峰. 营养液漂浮育苗棉苗根系适应水生环境的机理研究[J]. 中国农业科学, 2014, 47(17): 3372-3381.

ZHANG Hao, CHEN Jin-xiang, LIU Hai-he, ZHOU Zhong-hua, WANG Feng. Adaptation Mechanism to Aquatic Environment of Cotton Seedlings Roots in Floating Nursing Seedlings in Nutrient Water-Bed (FNSNW)[J]. Scientia Agricultura Sinica, 2014, 47(17): 3372-3381.

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