亚麻温敏雄性不育系的遗传、杂种优势和育性基因的RAPD标记研究
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摘要
亚麻(Linum usitatissimum L.)新型温敏雄性不育系的创建和研究,对于亚麻杂种优势利用具有十分重要的意义。本研究以新型亚麻温敏雄性不育系1S为研究对象,结合常规育种和分子生物学研究和分析方法,重点开展了雄性不育的遗传、杂种优势和育性基因的RAPD分子标记研究,取得的主要结论如下:
1.雄性不育的遗传
通过在自然条件下对不育系杂交后代F_1、F_2群体的育性鉴定,结果F_1代全部可育,F_2代育性发生分离,出现雄性可育和雄性不育株,表明该不育系的不育性受隐性核基因控制;对F_2群体育性分离数据的χ2测验表明,所有F_2调查群体育性分离均符合单基因遗传3:1的分离比,表明亚麻温敏雄性不育系的不育性受隐性单基因控制。
2.杂种优势表现
通过对6个杂交组合F_1、F_2代主要农艺性状和产量性状杂种优势的测定和分析,结果表明:不同杂交组合F_1、F_2代在株高和工艺长度上的平均优势均表现为负向超父本优势,而各产量性状上的平均优势均表现出很强的正向超父本优势;但与F_1代相比,F_2代平均优势明显下降。产量性状构成因素平均优势大小顺序为:单株产量>结实果数>千粒重>每果粒数。不同组合间F_1、F_2代杂种优势表现差异较大,其中1S×黑亚15号组合产量性状的超父本优势最强,F_1、F_2代均表现出很强的杂种优势。
3.亚麻基因组DNA最佳RAPD反应体系及程序的建立
通过优化组合,建立了适合本实验的亚麻基因组DNA最佳RAPD反应体系,25μl体系组成包括: 1U Tag酶、10×buffer( 200mM Tris-HCl、100mM (NH_4)_2SO_4、100 mM KCl、1% Triton X-100、pH 8.8) 2.5μl、2mM Mg~(2+)、150μM dNTPs、0.4μM random primer、30ng模板DNA。
优化反应程序为:94℃预变性5min;94℃、45s,37℃、60s,72℃、90s,共35个循环;最后72℃延伸10min。
A new thermo-sensitivity male-sterile line of Flax (Linum usitatissimum L.) were studied in this paper. With the research methods of traditional breeding and molecular biology, this work mainly focus on the analysis of heredity、heterosis and RAPD Markers of fertility gene for the new thermo-sensitivity male-sterile line 1S of Flax. The main results in this paper are as follow:
1. Heredity of the thermo-sensitivity male-sterile line of Flax
The identification of male fertility for F_1 and F_2 generations from six cross combinations under the natural conditions showed that all of F_1 populations were male fertile and in F_2 populations , there were fertile plant individuals and sterile ones, which indicated that the sterility of thermo-sensitivity male-sterile line of Flax was controlled by recessive nucleus genes. Through chi-square tests, the segregation ratios of fertile plants to sterile ones in observed F_2 segregation populations were coincided with the ratio 3:1 that is the segregation ratio of recessive single gene in F_2 generations.
2. Heterosis of the thermo-sensitivity male-sterile line of Flax
The heterosis of main agronomic and yield characters of F_1 and F_2 generations respectively from six cross combinations was analyzed. As far as the average heterosis of different cross combinations was concerned, both F_1 and F_2 genarations expressed negative over male-parent heterosis in plant height and technical length traits, however, positive over male-parent heterosis existed obviously in yield traits, including fruits per plant、seeds per fruit、1000-seed weight and seeds yield per plant. Compared with F_1 generations , the average heterosis of F_2 generations was lower.The order of heterosis is seed yield per plant>fruits per plant>1000-seed weight>seeds per fruit. The heterosis expression of F_1 and F_2 generations from different cross combinations was different. The heterosis of 1S×Heiya No.15 in yield traits was the largest among the six cross combinations not only in F_1 genarations but also in F_2 genarations.
1.雄性不育的遗传
通过在自然条件下对不育系杂交后代F_1、F_2群体的育性鉴定,结果F_1代全部可育,F_2代育性发生分离,出现雄性可育和雄性不育株,表明该不育系的不育性受隐性核基因控制;对F_2群体育性分离数据的χ2测验表明,所有F_2调查群体育性分离均符合单基因遗传3:1的分离比,表明亚麻温敏雄性不育系的不育性受隐性单基因控制。
2.杂种优势表现
通过对6个杂交组合F_1、F_2代主要农艺性状和产量性状杂种优势的测定和分析,结果表明:不同杂交组合F_1、F_2代在株高和工艺长度上的平均优势均表现为负向超父本优势,而各产量性状上的平均优势均表现出很强的正向超父本优势;但与F_1代相比,F_2代平均优势明显下降。产量性状构成因素平均优势大小顺序为:单株产量>结实果数>千粒重>每果粒数。不同组合间F_1、F_2代杂种优势表现差异较大,其中1S×黑亚15号组合产量性状的超父本优势最强,F_1、F_2代均表现出很强的杂种优势。
3.亚麻基因组DNA最佳RAPD反应体系及程序的建立
通过优化组合,建立了适合本实验的亚麻基因组DNA最佳RAPD反应体系,25μl体系组成包括: 1U Tag酶、10×buffer( 200mM Tris-HCl、100mM (NH_4)_2SO_4、100 mM KCl、1% Triton X-100、pH 8.8) 2.5μl、2mM Mg~(2+)、150μM dNTPs、0.4μM random primer、30ng模板DNA。
优化反应程序为:94℃预变性5min;94℃、45s,37℃、60s,72℃、90s,共35个循环;最后72℃延伸10min。
A new thermo-sensitivity male-sterile line of Flax (Linum usitatissimum L.) were studied in this paper. With the research methods of traditional breeding and molecular biology, this work mainly focus on the analysis of heredity、heterosis and RAPD Markers of fertility gene for the new thermo-sensitivity male-sterile line 1S of Flax. The main results in this paper are as follow:
1. Heredity of the thermo-sensitivity male-sterile line of Flax
The identification of male fertility for F_1 and F_2 generations from six cross combinations under the natural conditions showed that all of F_1 populations were male fertile and in F_2 populations , there were fertile plant individuals and sterile ones, which indicated that the sterility of thermo-sensitivity male-sterile line of Flax was controlled by recessive nucleus genes. Through chi-square tests, the segregation ratios of fertile plants to sterile ones in observed F_2 segregation populations were coincided with the ratio 3:1 that is the segregation ratio of recessive single gene in F_2 generations.
2. Heterosis of the thermo-sensitivity male-sterile line of Flax
The heterosis of main agronomic and yield characters of F_1 and F_2 generations respectively from six cross combinations was analyzed. As far as the average heterosis of different cross combinations was concerned, both F_1 and F_2 genarations expressed negative over male-parent heterosis in plant height and technical length traits, however, positive over male-parent heterosis existed obviously in yield traits, including fruits per plant、seeds per fruit、1000-seed weight and seeds yield per plant. Compared with F_1 generations , the average heterosis of F_2 generations was lower.The order of heterosis is seed yield per plant>fruits per plant>1000-seed weight>seeds per fruit. The heterosis expression of F_1 and F_2 generations from different cross combinations was different. The heterosis of 1S×Heiya No.15 in yield traits was the largest among the six cross combinations not only in F_1 genarations but also in F_2 genarations.
引文
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