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野生大豆和栽培大豆耐盐机理及遗传研究

Study on Mechanism and Inheritance of Salt Tolerance in Wild Soybean (Glycine Soja) and Cultivated Soybean (G.max)

【作者】 罗庆云

【导师】 刘友良;

【作者基本信息】 南京农业大学 , 植物学, 2003, 博士

【摘要】 评价、筛选并利用现有的野生种质资源来提高作物耐盐性,对开发利用盐碱土具有极其重要的意义。本研究以Lee68、南农1138-2、南农88-31、苏协1号、Jackson、中子黄豆乙等6个栽培大豆(Glycine max.L)品种和N23232、N23234、N23227、BB52(采集于山东省垦利县黄河入海口的盐碱滩涂)、JWS(采集于江苏省射阳县洋马乡的沿海滩涂)等5个野生大豆(G.soja.L)种群为材料,对各生育期耐盐性进行了鉴定;研究了NaCl胁迫对大豆茎叶表面腺体发育进程、分布密度及泌盐功能的影响;比较研究了NaCl胁迫下Cl~-和Na~+对野生和栽培大豆种子萌发、幼苗生长以及根系质子泵活性和叶片多胺含量的影响;研究了野生和栽培大豆对NaCl和Cl~-耐性的遗传规律。实验结果表明: NaCl胁迫下,野生大豆种子相对发芽指数为:BB52,JWS>N23232,N2234和N23227;出苗速率为:JWS>BB52>N23234,N23227>N23232;幼苗正常率为:JWS>BB52>N23227>N23232>N23234;全生育期耐盐性为:BB52=JWS>N23232。栽培大豆种子相对发芽指数为:Lee68,Jackson和南农88-31>中子黄豆乙和南农1138-2>苏协1号;出苗速率为:中子黄豆乙>Lee68>Jackson和苏协1号>南农88-31>南农1138-2;幼苗正常率为:南农1138-2>Jackson>中子黄豆乙和南农88-31>Lee68和苏协1号;苗期耐盐性为:Lee68和南农1138-2>南农88-31>苏协1号和Jackson>中子黄豆乙;全生育期耐盐性为:Lee68和南农1138-2>中子黄豆乙。上述结果表明,野生大豆各生育期耐盐性表现基本一致;栽培大豆苗期与全生育期耐盐性表现一致,而其他生育期与全生育期耐盐性表现不一致。 扫描电镜观察表明,球形“腺体”着生于植株茎杆及叶片远轴面(下表面)和近轴面(上表面)上,主要分布于远轴面尖部以及气孔和叶脉附近,与茎杆和叶片同步发育,并受NaCl胁迫的促进。但不为盐生野大豆BB52所特有。利用电子探针结合叶片漂洗研究表明,NaCl胁迫下,“腺体”表面矿质元素组成以Si为主,Na~+和Cl~-含量很低,无泌盐能力。 在种子萌发期和苗期,在等渗等浓度的单Cl~-、单Na~+和NaCl处理下,分别以相对发芽指数及植株受害程度为指标评价了NaCl胁迫下Na~+和Cl~-对野生和栽培大豆的胁迫作用,结果表明,栽培大豆对Cl~-敏感、对Na~+耐性强;野生大豆对Cl~-耐性强、对Na~+敏感。 在苗期,利用等渗的150mmol/L单Cl~-、单Na~+和NaCl处理并研究其对根系质膜罗庆云南京农业大学博士学位论文:野生大豆和栽培大豆耐盐机理及遗传研究H+一ATPase和液泡膜H十一PPas。、H气ATPase活性和叶片多胺组分的影响.结果表明,野生大豆根系质膜H九ATPase和液泡膜H+一PPase、H+一ATPase活性在胁迫ld时受激而明显高于对照,2d后下降到对照水平以下,其后又缓慢上升,BB52活性高于N23232;单Cl一处理下的液泡膜H气PPase活性明显高于单Na+及NaCI处理。 单Cl一处理下的野生大豆叶片高氛酸可溶性结合态多胺的相对含量显著高于单Na+和NaCI处理;同一处理下,BB52叶片高氛酸可溶性结合态多胺相对含量较N23232高。单Na+处理下的耐盐栽培大豆叶片高氛酸可溶性结合态多胺相对含量显著高于单Cl一和NaCI处理。因此,游离态和高氛酸不溶性结合态多胺向高氛酸可溶性结合态多胺的转化可能与大豆耐盐性增强有关。 以野生和栽培大豆及栽培大豆间配制的5个杂交组合的PI、PZ、FI、F:和凡3世代为材料,牙.J用多世代联合分离分析方法分析了野生和栽培大豆对Cl‘和NaCI耐性的遗传规律。实验结果表明,栽培大豆的Cl一耐性受一对主基因控制,同时存在微效基因效应;野生大豆的Cl一耐性受与栽培大豆不同的基因控制,其与栽培大豆杂交后代的a一耐性高于栽培大豆间的杂交后代。Lee68xN23227组合对NaCI的耐性遗传符合一对加性主基因+加胜一显性多基因模型.中子黄豆乙xBB52和栽培大豆间杂交组合南农88一31‘Jackson和南农1 1 38一2x南农88一31对NaCI的耐性遗传符合加性一显性一上位性多基因遗传模型。

【Abstract】 Evaluation, selection and utilization of wild germ plasm to enhance salt tolerance of plants by breeding is very important to the exploitation of saline soil. 6 species of soybean cultivars (Glycine max L.)-Lee68, Nannong 1138-2, Nannong 88-31, No.1 of Suxie, Jackson and Zhongzi Huangdou-Yi and 5 populations of wild soybean (G soja L.)-N23232, N23234, N23227, BB52 (collected from the setuary of the Yellow Rive in Kenli County, Shandong Province) and JWS (collected from the beach in Sheyang County, Jiangsu Province) were used as plant materials in this research. At the first, evaluated for their ability of salt tolerance at various stages of growth and development were evaluated; At the second, effects of NaCl stress on the development and salt secretion of glands on surface of stems and leaves in G max as well as G soja were investigated. At the third, stress effects of Na+ and Cl- caused by NaCl on seed germination, seedlings growth, H+-ATPase activities of plasma membrane, H+-ATPase and H+-PPase activities of tonoplast of the roots and contents of polyamines of the leaves in G max and G soja were compared. Finally, inheritance of NaCl and Cl" tolerance in G max and G soja were investigated. Result showed that:Under NaCl stress, among the 5 G soja populations, seed relative germination index were BB52, JWS>N23232, N23234 and N23227, seed emergence rate were JWS>BB52> N23227>N23232>N23234, salt tolerance at whole development stage were JWS, BB52>N23232. Among the 6 G max cultivars, under NaCl stress, seed relative germination index were Lee68, Jackson, Nannong 88-31>Zhongzi Huangdou-Yi, Nannongll38-2> No.1 of Suxie, seed emergence rate were Zhongzi Huangdou-Yi> Lee68> Jackson, No.l of Suxie>Nannong88-31> Nannongl 138-2, salt tolerance of seedlings were Lee68, Nannong 1138-2>Nannong 88-31> No.l of Suxie, Jackson> Zhongzi Huangdou-Yi, salt tolerance at whole development stage were Lee68, Nannong 1138-2> Zhongzi Huangdou-Yi. These results indicated that salt tolerance of G soja populations at various stages were performance consistence but inconformity for G max cultivars; and the salt tolerance atseedling stage were performance consistence with those of the whole development stage for G max cultivars.The results observed with scanning electron microscope showed that, the globe "glands" were born in surface of stems and the adaxial/abaxial surface of leaves, and mainly attached to the epidermal cells in abaxial surface of leaf apex, near and on the main/later vein and guard cells of stoma. And their development progress was synchronizing with the leaf and stem it attached and their density was promoted by NaCl stress. By the way, the globe "glands" were not monopolist by the seedlings of BB52, and were founded in seedlings of other populations of G. soja and cultivar of G max. The results studied with electro-probe (X-ray microanalysis) and leaf rinsing showed that, under NaCl stress, chemical element constitute in the globe "gland" surface regarding Si as the lord, contents of Na+ and Cl- were very low. The globe "gland" couldn’t secrete salt.At seedling emergence stage and seedling stage, the relative germination index and salt injury suffered under isoosmotic stress of Cl-, Na+ and NaCl at equal concentration were used to evaluate and compare the stress effects of Cl" and Na+ on G max cultivars and G soja populations under NaCl stress. Results showed that, cultivars of G max were more sensitive to stress effect from Cl" than that from Na+; however, populations of G. soja were more sensitive to stress effect from Na^ than that from Cl".In the period of seedling, content of polyamines in leaves as well as activities of I-T-ATPase of plasma membrane and activities of H+-PPase and H+-ATPase of toloplast in roots of G soja populations were mensurated under isoosmotic stress of 150 mmol/ L Cl-, Na+ and NaCl. Results showed that, the activities of plasma membrane H+-ATPase, toloplast H+-PPase and H+-ATPase increased after stress for 1 day; after stress for 2 days, their activities decreased to the levels

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