利用EMS诱变构建甘蓝型油菜突变体库的初步研究
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摘要
突变体在农作物品种改良、功能基因研究等方面具有十分重要的意义。本研究采用4种浓度EMS处理甘蓝型油菜NJ7982种子,选取其中诱变效果好的处理(0.4%EMS)所衍生的群体,作本研究的对象。通过对诱变后代的苗期、开花期、成熟期的表型观察,对表型性状突变体进行筛选和鉴定；还对突变处理群体进行了种子脂肪酸含量筛选和除草剂筛选。主要结果如下：
1.表型突变性状：对M2群体内各植株的形态学观察,获得了包含叶、花器、株型、角果、种皮的表型性状的突变体,共8789株,总的突变表型频率达18.51%。子叶性状突变体包括3子叶、子叶黄化等,占群体的0.22%；叶片性状突变体包括有黄化叶、白化叶、紫色叶、上卷叶、下卷叶等,占群体0.74%;花器性状突变体包括紫色花蕾、死蕾、3花瓣、6花瓣、白色瓣、黄白双色镶嵌花瓣、花瓣皱缩、完全不育、微粉株等,占群体9.38%;株型性状突变体包括矮杆、粗茎、多分枝、紧凑型、丛生、褐茎、紫茎、早衰等,占群体4.98%;角果性状突变体包括粗角、长角、弯角、紫角等,占2.79%；另外,在收获群体内筛选获得了50份黄籽材料,占收获群体0.40%。这些多样性的表型突变材料,为甘蓝型油菜功能基因研究及种质创新提供基础性材料。
2.高油酸种质：通过种子脂肪酸含量测定,从M2群体内筛选获得了7份油酸含量大于73%的种质,平均油酸含量达75.18%±1.69%,与野生型的(66.65%)相比较提高了8.53个百分点。这些高油酸材料在油菜脂肪酸改良中将具有十分重要的应用价值。
3.抗草甘膦变异株：对M2代105 263株幼苗进行草甘膦喷雾,处理后仅5株幼苗成活下来,幼苗成活率达到0.004%,并认为此5株幼苗为抗除草剂材料。M3代,重复性鉴定的结果表明,变异材料对致死浓度的草甘膦仍具有忍耐力,这为抗除草剂油菜品种的培育提供新材料。
Mutants have been playing important role in crop genetic improvement and functional genomics researches.The present research constructed a mutant library derived from an EMS treatment which had the best inducement effect in 4 EMS concetrations(0%,0.2%, 0.4%,0.6%) treatment of canola line NJ7982 (Brassica napus L) from Nanjing Agricultural University. Through morphorlogical observation, seed quality determination and herbicide resistance evaluation in mutation progenies, phenotype mutants have been recorded and collected, high oleic acid content mutants and herbicide-resistant plants have also been obtained. Main results are as follows.
1. Various phenotypic mutants have been obtained. In M2 population,8789 plants with phenotypic mutative traits which were devided into classes such as leaf, flower, plant type, silique and seed coat trait mutants, were observed, accounting for 18.51% of the M2 plants. Cotyledon trait mutants covered tricotyledon. yellowish cotyledon, etc. accounting 0.22% in the M2 population. Leaf trait mutants such as yellowish leaf, albino leaf, purple leaf, up-curly leaf, down-curly leaf, etc, were observed with mutation frequency of 0.74%. Flower trait mutants manifested as purple buds, bud-deads, three petals, six petals, white petal, white-mosaic petal, wrinkled-petal, complete male sterility, few powder, etc, were of 9.38% in the M2 population. Plant type trait mutants included dwarf plant, thick stem, excessive branches, tight type, tufty branches, brown stem, purple stem, premature, etc, accounting for 4.98%. Silique trait mutants accounting for 2.79% included thick-walled silique, long silique. curly silique, purple silique, etc. Fifty yellow-seeded mutants were also obtained, accounting 0.40% in harvested population. All these phenotypic mutation materials were saved in germplasm bank, will be used to benefit rapeseed functional genomics research and variety improvement.
2. Germplasm with high oleic acid content was obtained. By determinating seed fatty acid content, screening results showed that seven germplasms with more than 73% oleic acid content in seed oil were obtained from M2 plants with average oleic acid content of 75.18%, higher than 66.65% of its wild type by 8.53%, will be very useful in fatty acid improvement of Brassica napus.
3. Herbicide-resistant plants were obtained. By spraying glyphosate, five mutants were alive after treatment from 105 263 seedlings, and the survival rate of seeding reached 0.004% in the population. Evaluation of glyphosate with carious concentrations in M3 showed that the mutation materials were also resistant to herbicide, and provide new materials for herbicide-resistant variety breeding.
1.表型突变性状：对M2群体内各植株的形态学观察,获得了包含叶、花器、株型、角果、种皮的表型性状的突变体,共8789株,总的突变表型频率达18.51%。子叶性状突变体包括3子叶、子叶黄化等,占群体的0.22%；叶片性状突变体包括有黄化叶、白化叶、紫色叶、上卷叶、下卷叶等,占群体0.74%;花器性状突变体包括紫色花蕾、死蕾、3花瓣、6花瓣、白色瓣、黄白双色镶嵌花瓣、花瓣皱缩、完全不育、微粉株等,占群体9.38%;株型性状突变体包括矮杆、粗茎、多分枝、紧凑型、丛生、褐茎、紫茎、早衰等,占群体4.98%;角果性状突变体包括粗角、长角、弯角、紫角等,占2.79%；另外,在收获群体内筛选获得了50份黄籽材料,占收获群体0.40%。这些多样性的表型突变材料,为甘蓝型油菜功能基因研究及种质创新提供基础性材料。
2.高油酸种质：通过种子脂肪酸含量测定,从M2群体内筛选获得了7份油酸含量大于73%的种质,平均油酸含量达75.18%±1.69%,与野生型的(66.65%)相比较提高了8.53个百分点。这些高油酸材料在油菜脂肪酸改良中将具有十分重要的应用价值。
3.抗草甘膦变异株：对M2代105 263株幼苗进行草甘膦喷雾,处理后仅5株幼苗成活下来,幼苗成活率达到0.004%,并认为此5株幼苗为抗除草剂材料。M3代,重复性鉴定的结果表明,变异材料对致死浓度的草甘膦仍具有忍耐力,这为抗除草剂油菜品种的培育提供新材料。
Mutants have been playing important role in crop genetic improvement and functional genomics researches.The present research constructed a mutant library derived from an EMS treatment which had the best inducement effect in 4 EMS concetrations(0%,0.2%, 0.4%,0.6%) treatment of canola line NJ7982 (Brassica napus L) from Nanjing Agricultural University. Through morphorlogical observation, seed quality determination and herbicide resistance evaluation in mutation progenies, phenotype mutants have been recorded and collected, high oleic acid content mutants and herbicide-resistant plants have also been obtained. Main results are as follows.
1. Various phenotypic mutants have been obtained. In M2 population,8789 plants with phenotypic mutative traits which were devided into classes such as leaf, flower, plant type, silique and seed coat trait mutants, were observed, accounting for 18.51% of the M2 plants. Cotyledon trait mutants covered tricotyledon. yellowish cotyledon, etc. accounting 0.22% in the M2 population. Leaf trait mutants such as yellowish leaf, albino leaf, purple leaf, up-curly leaf, down-curly leaf, etc, were observed with mutation frequency of 0.74%. Flower trait mutants manifested as purple buds, bud-deads, three petals, six petals, white petal, white-mosaic petal, wrinkled-petal, complete male sterility, few powder, etc, were of 9.38% in the M2 population. Plant type trait mutants included dwarf plant, thick stem, excessive branches, tight type, tufty branches, brown stem, purple stem, premature, etc, accounting for 4.98%. Silique trait mutants accounting for 2.79% included thick-walled silique, long silique. curly silique, purple silique, etc. Fifty yellow-seeded mutants were also obtained, accounting 0.40% in harvested population. All these phenotypic mutation materials were saved in germplasm bank, will be used to benefit rapeseed functional genomics research and variety improvement.
2. Germplasm with high oleic acid content was obtained. By determinating seed fatty acid content, screening results showed that seven germplasms with more than 73% oleic acid content in seed oil were obtained from M2 plants with average oleic acid content of 75.18%, higher than 66.65% of its wild type by 8.53%, will be very useful in fatty acid improvement of Brassica napus.
3. Herbicide-resistant plants were obtained. By spraying glyphosate, five mutants were alive after treatment from 105 263 seedlings, and the survival rate of seeding reached 0.004% in the population. Evaluation of glyphosate with carious concentrations in M3 showed that the mutation materials were also resistant to herbicide, and provide new materials for herbicide-resistant variety breeding.
引文
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