本文選題：野生大豆　切入點：多樣性分析　出處：《山東師范大學》2017年碩士論文

【摘要】：人們在耕作過程中的不合理灌溉、施肥、自然災害等因素使得全球的土地鹽堿化日趨嚴重。大豆是我國主要的油料作物,但隨著育種進程中的不斷選擇,栽培大豆遺傳多樣性日趨狹窄。野生大豆是栽培大豆的野生近緣種,并且含有很多優(yōu)異基因,例如:耐鹽、抗旱、耐寒、抗蟲、多莢等,尤其在耐鹽方面比栽培大豆有著顯著的優(yōu)勢。本研究以327份野生大豆種質資源為材料,通過表型性狀及SSR分析,篩選并建立野生大豆多樣性群體,對該群體進行表型性狀及抗逆性多樣性分析。并對其中耐鹽及鹽敏感極端材料進行耐鹽生理生化分析、研究耐鹽相關基因的表達情況,解析野生大豆耐鹽機理。利用大豆發(fā)狀根體系,通過耐鹽相關基因的過表達及CRISPR-cas9技術突變目的基因,研究耐鹽相關基因功能,為大豆耐鹽育種提供重要信息。主要研究結果如下:1.對327份野生大豆資源的主莖類型、葉形、葉長、葉寬、葉柄長度、莢長、莢寬、花色等農藝性狀進行調查,發(fā)現這些性狀存在豐富的多樣性,但有些材料相似性很高。利用課題組以前篩選出的具有較高多態(tài)性的10對SSR引物對327份野生大豆進行SSR分析,去除表型及SSR標記一致的材料,最終建立了101份野生大豆的多樣性群體。2.對101份野生大豆多樣性群體進行遺傳多樣分析發(fā)現,主莖類型、葉長、葉寬、葉柄長度、葉面積、莢長、莢寬、單株籽粒重、苗期株高、干重、耐鹽性等方面均表現出了豐富的多樣性,變異系數0.18～0.79,多樣性指數為1.73～2.05。3.對耐鹽材料ST-335和鹽敏感材料SS-736進行鹽脅迫下生理生化分析,發(fā)現鹽脅迫條件下,與鹽敏感品種SS-736相比,耐鹽品種ST-335的葉片相對含水率高,可溶性糖含量高;SOD、POD和CAT活性高;Na+/K+低;且KOR,NHX和NSCC基因表達量低,而SKOR和SOS1基因表達量高。可能是ST-335具有較高的抗離子毒害、滲透脅迫及抗氧化脅迫的能力,且離子轉運相關的一些基因通過協同作用控制Na+及K+的吸收和外排,最終決定ST-335具有相對較高的耐鹽能力。4.在大豆發(fā)狀根中過表達SKOR和SOS1基因,提高了大豆復合體的耐鹽能力;而過表達NSCC基因,提高了大豆復合體對高鹽脅迫的敏感性。在野生大豆發(fā)狀根中利用CRISPR/Cas9體系成功誘導NSCC基因發(fā)生突變,NSCC發(fā)生突變的發(fā)狀根中的Na+/K+比例顯著低于野生型。進一步說明這些離子通道相關基因的變化對野生大豆耐鹽能力產生影響。該方法也為野生大豆耐鹽相關基因功能研究提供重要的理論基礎。
[Abstract]:People in the farming process unreasonable irrigation, fertilization, natural disasters and other factors make the global land salinization has become increasingly serious. Soybean is the main oil crop in China, but with the continuous selection in the process of soybean genetic diversity is becoming more and more narrow. Wild soybean is the wild relatives of cultivated soybean, and contains a lot of excellent genes, such as salt tolerance, drought resistance, cold resistance, insect resistance, multi pod, especially in salt tolerance than cultivated soybean has the remarkable superiority. In this study, 327 wild soybean germplasm, through the analysis of phenotypic traits and SSR screening, and the establishment of wild soybean population diversity, analyze the Phenotype Traits and resistance diversity of the group. And the salt tolerant and salt sensitive materials for extreme salt tolerance physiological and biochemical analysis, the expression of genes related to salt tolerance, salt tolerance mechanism of wild soybean using. Soybean hairy root system by mutation gene expression and CRISPR-cas9 gene related to salt tolerance, salt tolerance related gene function research, provide important information for salt tolerance in soybean breeding. The main results are as follows: 1. of the 327 wild soybean resources stem type, leaf shape, leaf length, leaf width, petiole length. Pod length, pod width, color and other agronomic traits were investigated and found that the rich diversity of these traits, but some material similarity is very high. The research group previously screened with high polymorphisms of 10 SSR primers of 327 wild soybean by SSR analysis, SSR marker phenotype and removal consistent material. The eventual establishment of 101 wild soybean population diversity.2. of 101 wild soybean population diversity of genetic diversity analysis showed that the main stem type, leaf length, leaf width, petiole length, leaf area, pod length, pod width, grain weight per plant, seedling High, dry weight, salt tolerance has showed the rich diversity, the coefficient of variation was 0.18 ~ 0.79, the diversity index was 1.73 ~ 2.05.3. of salt tolerant and salt sensitive material ST-335 material SS-736 under Salt Stress on physiological and biochemical analysis, found that under salt stress, compared with salt sensitive strains of SS-736. Leaves of salt tolerant cultivar ST-335 relatively high moisture content, high content of soluble sugar; SOD, POD and CAT activity; Na+/K+ and KOR, is low; the expression of NHX and NSCC gene is low, and the expression of SKOR and SOS1 gene may be high. ST-335 has high anti permeability ability ion toxicity, oxidative stress and stress and, some genes related ion transport through synergistic absorption and control of Na+ and the role of K+, ST-335 finally decided to have a relatively high salt tolerance of.4. over expression of SKOR and SOS1 genes in soybean hairy root, improve soybean complex salt tolerance; and The expression of NSCC gene increased soybean complex sensitivity to high salt stress induced by CRISPR/Cas9 system. The NSCC gene mutation in wild soybean hairy roots, NSCC mutation of hairy root in the proportion of Na+/K+ was significantly lower than that of wild type. Further changes of the ion channel genes related to salt tolerance of wild soybean effect. The method also provides important theoretical basis for research on wild soybean salt tolerance related gene function.

【學位授予單位】：山東師范大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：S565.1

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