马铃薯抗旱资源的筛选及抗旱相关基因的鉴定

发布时间：2018-06-23 15:54

本文选题：马铃薯 + 抗旱资源筛选　；参考：《东北农业大学》2017年硕士论文

【摘要】：干旱对农业生产的危害日趋严重,抗旱作物品种的种植是应对干旱的有效途径之一,因此作物抗旱资源的筛选对其适应干旱环境具有重大意义。我国马铃薯主产区的大部分区域为水资源匮乏的干旱贫瘠地区,随着全球气候变暖,干旱地区的面积逐年扩大,因此抗旱资源的筛选与育种利用、以及抗旱品种在生产上的推广应用,将对提高干旱地区马铃薯的产量发挥重要作用;有效发掘关键调控基因,对深入研究马铃薯干旱胁迫应答的分子机理具有重要意义。本试验对210份抗旱育种后代材料进行PEG-6000模拟干旱和抗旱棚种植,利用几种抗旱生理指标综合评价的方法,筛选出了抗旱性好的材料;利用从已知抗旱品种中发掘的与抗旱相关的基因,对抗旱性好且综合表现优良的材料开展转录水平检测。为马铃薯抗旱种质资源的筛选、抗旱性快速评价体系的建立和抗旱育种工作的开展奠定了基础。主要研究结果如下:(1)利用20%PEG-6000溶液胁迫离体叶片法对210份抗旱育种后代材料进行抗旱性初步评价,根据萎蔫程度、抗旱生理指标,并结合产量,初步筛选出17份抗旱材料,建立了一套快速有效筛选及评价马铃薯抗旱种质资源的的方法。(2)采用抗旱棚盆栽控水试验对入选的17份材料进行抗旱性综合评价,测定农艺性状、抗旱生理指标、光合参数及产量,利用隶属函数分析法鉴定出高度抗旱材料6份,其中ND23、ND20的抗旱性强于定薯1号;ZD33、ZD29的抗旱性强于克新1号,但弱于定薯1号;ND2的抗旱性强于冀张薯8号,但弱于克新1号,ND22的抗旱性强于东农311,但弱于冀张薯8号。(3)针对8个在干旱诱导下蛋白呈现上调表达的相关基因,进一步进行抗旱功能的鉴定。利用20%PEG-6000溶液胁迫处理已知抗旱性有差异的品种克新1号、冀张薯8号和费乌瑞它的脱毒试管苗,进行Real-time PCR(q RT-PCR)检测,结果表明:有5个基因可能对抗旱调节起到关键作用,分别为热休克蛋白STI基因(HSPSL)、ATP合酶基因(ATPSM)、甘氨酸富集蛋白基因(GRP2L)、叶绿体50S核糖体蛋白基因(50RP4L)、马铃薯叶绿体核酮糖-5-磷酸3-差向异构酶基因(P5P3E)。(4)对6份高度抗旱材料进行5个抗旱相关基因的q RT-PCR检测,结果表明:ND23、ND22、ZD33、ZD29、ND2中,5个抗旱相关基因上均显著上调表达;ND20(除GRP2L基因)有4个抗旱相关基因显著上调表达。HSPSL基因、GRP2L基因上调表达时间较早,最大上调表达时间是6 h、12 h和24 h;ATPSM基因、50RP4L基因在干旱胁迫的前期上调表达,上调表达时间是6 h和12 h;P5P3E基因分别在干旱胁迫的早、晚期上调表达,上调表达时间是6 h和48 h。材料的抗旱性在转录水平上得到验证,基因在高度抗旱材料中上调表达。
[Abstract]:Drought damage to agricultural production is becoming more and more serious. Planting drought-resistant crop varieties is one of the effective ways to cope with drought. Therefore, the screening of drought-resistant crop resources is of great significance to their adaptation to drought environment. The majority of potato production areas in China are arid and barren areas with scarce water resources. With the global warming, the area of arid areas is expanding year by year, so the selection and breeding utilization of drought-resistant resources. The popularization and application of drought-resistant varieties in production will play an important role in improving the yield of potato in arid areas, and it is of great significance to explore the key regulatory genes and to further study the molecular mechanism of drought stress response in potato. In this experiment, 210 progenies of drought resistance breeding were planted in simulated drought and drought resistant shed with PEG-6000. The materials with good drought resistance were screened by using several comprehensive evaluation methods of physiological indexes of drought resistance. The transcriptional level of drought-resistant materials with good drought resistance and comprehensive performance was detected by using genes related to drought resistance which were extracted from known drought-resistant varieties. It lays a foundation for the screening of drought resistant germplasm resources, the establishment of rapid evaluation system for drought resistance and the development of drought resistance breeding. The main results are as follows: (1) the drought resistance of 210 drought resistant breeding progenies was preliminarily evaluated by using 20 PEG-6000 solution stress in vitro. According to wilting degree, drought resistance physiological index and yield, 17 drought resistant materials were preliminarily selected. A set of rapid and effective methods for screening and evaluating drought resistant germplasm resources of potato were established. (2) the drought resistance of 17 selected materials was evaluated by pot water control experiment in drought-resistant greenhouse, and agronomic characters and physiological indexes of drought resistance were determined. Photosynthetic parameters and yield were identified by membership function analysis. The drought resistance of ND23 Nd20 was stronger than that of Dingshu 1, ZD33, ZD29 and Kexin 1, but weaker than that of Dingshu 1 and Dingshu 1, but less than that of Dingshu 1 and Jizhangshu 8, respectively, and the drought resistance of ND20 was stronger than that of Dingshu 1, ZD33 and ZD29, but weaker than that of Dingshu 1 and Nd2. But the drought resistance of Nd22 was weaker than that of Dongnong 311, but weaker than that of Jizhangshu 8. (3) the drought resistance function of 8 genes which showed up-regulated expression under drought induction was further identified. Using 20 PEG-6000 solution stress to treat virulent plantlets of known drought-resistant varieties Kexin 1, Ji Zhangshu 8 and Fiurita, and detect them by Real-time PCR (Q RT-PCR). The results showed that 5 genes may play a key role in drought resistance regulation. Heat shock protein STI gene (HSPSl), glycine rich protein gene (GRP2L), chloroplast 50S ribosomal protein gene (50RP4L) and chloroplast ketoglycose-5-phosphate 3-differential isomerase gene (P5P3E). (4) were used to treat 6 drought resistant materials. Five drought-related genes were detected by qRT-PCR. The results showed that 4 of the 5 drought related genes (except GRP2L gene) were significantly up-regulated in 4 of the 5 drought related genes (except GRP2L gene). The up-regulation of GRP2L gene was earlier than that of HSPSL gene. The maximum up-regulated expression time was 6 h, 12 h and 24 h, respectively, and the up-regulation time was 6 h and 12 h, respectively, and the up-regulation time was 6 h and 48 h, respectively. The drought resistance of the materials was verified at the transcriptional level, and the gene expression was up-regulated in the highly drought-resistant materials.
【学位授予单位】：东北农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S532

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