西南地区玉米地方品种B染色体多态性分析

doi:10.3864/j.issn.0578-1752.2015.14.002

摘要/Abstract

摘要： 【目的】分析B染色体形态及数量多态性，从细胞水平评价西南地区玉米地方品种的遗传多样性，为中国玉米引进途径提供佐证。【方法】以来自西南地区四川、重庆、云南和贵州4省（市）的30个玉米地方品种群体为材料，各品种群体取50个单株，各单株统计镜检10个细胞，共统计镜检15 000（30 × 50 × 10）个细胞，在光学显微镜下进行B染色体细胞学鉴定；各品种群体取10个单株，制作10张染色体制片，采用改良的Giemsa染色法进行染色体C-带显带分析。【结果】细胞学鉴定结果表明，除10对常染色体外，西南地区玉米地方品种还存在3类形态的B染色体：中间着丝点B染色体（B_M）、近端着丝点B染色体（B_ST）和呈点状的微小B染色体（B_S），B_M形态上与玉米的第1染色体相似，B_ST与B_M大小相近，B_S呈圆点状，无可见着丝点。在来自四川的7个玉米地方品种群体中检测到B_M、B_ST和B_S3类B染色体，在重庆玉米地方品种群体中检测到B_M和B_ST、B_ST和B_S存在于云南玉米地方品种群体，而在来自贵州的9个玉米地方品种群体中仅检测到1类B染色体（B_ST）。B_M、B_ST和B_S的平均长度分别为2.82、2.78和0.9 µm，与第1常染色体比较，它们的平均相对长度分别为43%、45%和15%，相对长度变幅分别为34%—52%、32%—58%和13%—18%，B_M、B_ST和B_S的平均臂比分别为1.19、1.98和1.00。在供试的30个品种群体中有18个含B染色体，分别有7个、5个、4个和2个来自四川、重庆、云南和贵州，在含B染色体的18个群体中有421个单株含B染色体，421个单株中共存在487条B染色体，具有B染色体植株的频率变幅为0—90%，平均频率为29.6%，四川玉米地方品种在群体和个体水平上表现较高的B染色体频率。细胞内B染色体数最多2个，绝大多数细胞不含B染色体，0B类、1B类和2B类细胞的平均频率分别为97.04%、2.81%和0.15%。B_ST是玉米地方品种B染色体的主要类型，占B染色体总数的比率为67%，其次是B_M（19%），B_S比率最低（14%），表明B_ST是原始的B染色体，B_M和B_S是B_ST的变异类型，B_M和B_S在较小的时间和空间尺度上形成与演化。Giemsa染色显示，B_M和B_ST富含GC、高度异染色质化，在染色体结构上二者共享部分同源DNA序列，形态上与A染色体区别明显。0B类、1B类和2B类细胞A染色体平均C-带数变幅分别为11.0—20.6、10.8—19.4和10.5—18.6，玉米地方品种B染色体数与A染色体C-带数负相关。【结论】西南地区玉米地方品种在细胞水平上表现较高的遗传多样性，其B染色体具有形态和数量多态性，由B染色体分布所决定的西南地区玉米地方品种的地理中心是四川省。

关键词: 玉米, B染色体, 多态性, 西南地区

Abstract: 【Objective】 The objective of this study is to analyze B chromosome polymorphisms in maize landrace populations from Southwest China, to evaluate their genetic diversity at cellular level and provide evidence for the introduction path of maize in China. 【Method】 Thirty maize landrace populations from Sichuan Province, Chongqing city, Yunan Province, and Guizhou Province in southwestern China were used. Fifty individuals per landrace population were sampled, and ten cells from a single plant were scored for the presence of Bs under a microscope. A total of 15 000 cells (30 × 50 × 10) in the mitotic metaphase were analyzed. Ten slides, from ten individuals per landrace population, were C-banded using conventional Giemsa staining with modifications.【Result】 Besides 10 pairs of standard chromosomes, the landrace populations contain a metacentric B chromosome (B_M) morphologically similar to the first chromosome of the karyotype, a subtelocentric B (B_ST) identical in size to B_M, and a spot-shape microchromosome (B_S) without centromere. Seven maize landrace populations from Sichuan possessed three forms of Bs. In the populations of maize landraces from Chongqing both B_M and B_ST forms were found, whereas B_ST and B_S were identified in the populations of maize landraces from Yunnan. Nine populations from Guizhou were the only one (B_ST) where B_M and B_S were exclusive. The mean length of B_M, B_ST, and B_S was 2.82 µm, 2.78 µm, and 0.9 µm, respectively. In comparison with the metacentric pair of the standard karyotype, measurements conducted on the cells with Bs estimated a relative length of 34%-52% for the B_M, 32%-58% for B_ST, and 13%-18% for the B_S, and relative lengths for B_M, B_ST, and B_S were on the average 43%, 45%, and 15%, respectively. The mean arm ratios for B_M, B_ST, and B_S were 1.19, 1.98, and 1.00, respectively. Of the eighteen populations with B chromosomes, seven, five, four, and two populations originated geographically from Sichuan, Chongqing, Yunnan, and Guizhou, respectively. Eighteen of the thirty populations sampled exhibited numerical polymorphisms and a total of 487 Bs were identified in 421 individuals. The frequency of B chromosomes per individual varied from 0 to 90% with an average of 29.6%. The populations from Sichuan possess a higher frequency of B chromosomes in both population and individual levels. The highest number of B chromosomes in a cell was 2 with the predominant dose being 0. The mean frequency of B chromosomes for the 0B, 1B, and 2B cell were 97.04%, 2.81%, and 0.15%, respectively. B_ST is the predominant form accounting for 67% of the Bs in the populations, followed in turn by B_M and B_S corresponding to 19% and 14%, respectively. It might be hypothesized that B_ST represented an ancestor B chromosome in maize, whereas the B_ST and B_S were variants of the B_M, implying that the evoluation of B_M and B_S have occurred in a smaller time and space scale. The Giemsa staining exhibited that the B_M and B_ST forms, easily distinguished from A chromosomes, shared some homologous DNA sequences and were both heterochromatic as well as GC-rich. The mean C-band number varied from 11.0 to 20.6 in the 0B cell, 10.8 to 19.4 in the 1B cell, and 10.5 to 18.6 in the 2B cell, indicating a negative correlation between B chromosomes and C-bands in the A chromosome set. 【Conclusion】 Cytological observations revealed that maize landrace populations from southwest China had a higher level of genetic diversity. For the B chromosome in them, there existed morphological and numerical polymorphisms. Sichuan is the geographical centre of maize landraces in this region based on geographical distributions of the B chromosome. This supported the theory on a maize introduction path in China.

Key words: maize, the B chromosome, polymorphisms, southwest China

姚启伦，陈发波，刘红芳，方平，赵彩芳. 西南地区玉米地方品种B染色体多态性分析[J]. 中国农业科学, 2015, 48(14): 2697-2704.

YAO Qi-lun, CHEN Fa-bo, LIU Hong-fang, FANG Ping, ZHAO Cai-fang. B Chromosome Polymorphisms in Maize (Zea mays L.) Landrace Populations from Southwest China[J]. Scientia Agricultura Sinica, 2015, 48(14): 2697-2704.

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