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构建番茄突变体库对于丰富遗传变异、发掘基因资源具有重要意义。利用1%的甲基磺酸乙酯(EMS)处理‘里格尔87-5’加工番茄种子12 h,对M_2代材料进行农艺学性状与生物学性状鉴定,对部分M_3代材料进行遗传稳定性验证。在M_2代筛出典型突变体,包括早衰、矮化、圆果、低抗坏血酸、黑斑、绿茎、簇生等突变单株。对240个M_2代家系中共1 023个单株全生育期田间表型进行观察鉴定和果实品质测定,共发现90个变异单株,其中13个突变单株同时出现两个或两个以上变异性状,共50个变异性状,总的单株变异频率(突变株/总株数)为8.80%。叶、茎、花、果实和植株发生的变异株数分别为27、7、17、56和21,性状变异频率分别为2.64%、0.68%、1.66%、5.47%和2.05%。其中矮化突变体分子鉴定表明,curl-3基因外显子中C变成T,氨基酸由L变成F,喷施油菜素内酯不能恢复其正常生长,表明curl-3突变导致油菜素内酯信号传导受阻。curl-3突变体叶片与果实中抗坏血酸含量下降,并伴随着叶片中氧化态与还原态抗坏血酸比值增加,这可能与抗坏血酸氧化酶基因AO表达量和AO酶活性增强有关。
Constructing a tomato mutant library is of great significance for enriching genetic variation and exploring genetic resources. Agrochemical traits and biological traits of M_2 generation materials were determined by using 1% ethyl methylsulfate (EMS) for processing seed of ’Ligel 87-5’ for 12 h, and genetic stability of some M_3 generation materials Sexual verification. Typical mutants were screened on M_2, including mutant individuals such as premature aging, dwarfing, round fruit, low ascorbic acid, dark spots, green stems and tufted plants. The field phenotypes of 1 023 whole plantlets in 240 M_2 generation families were observed and identified and the fruit quality was measured. A total of 90 mutant individuals were found, in which 13 mutant plants showed two or more variation traits at the same time , A total of 50 variation traits, the total variation frequency of single plant (mutant / total number of plants) was 8.80%. The number of variation of leaves, stems, flowers, fruits and plants were 27, 7, 17, 56 and 21, respectively. The frequency of trait variation was 2.64%, 0.68%, 1.66%, 5.47% and 2.05%, respectively. The molecular identification of the dwarf mutant showed that C in the exon of the curl-3 gene became T and the amino acid changed from L to F, and the brassinolide could not restore its normal growth, indicating that the mutation of curl-3 leads to intra-brassinin Ester signaling is blocked. The content of ascorbic acid in leaves and fruits of curl-3 mutant decreased with the increase of the ratio of oxidation state to reduced ascorbic acid in leaves, which might be related to the increase of AO expression level and AO activity of ascorbate oxidase gene.