基于顺序GISH-FISH花生栽培种的染色体分析

doi:10.3864/j.issn.0578-1752.2015.09.19

Abstract

Abstract: 【Objective】The cytogenetic study of peanut has been hindered mainly by the small chromosomes and limited cytological markers of peanut. In order to reveal the correspondence between cultivated peanut (Arachis hypogaea L., 2n=4x=40, AABB) and wild donor parents chromosomes and to characterize the chromosome structure variants, the objective of this study is to establish a highly distinguishable karyotype of A and B genome chromosomes for improving accuracy of chromosome identification. 【Method】Using the total genomic DNA of the two possible donors (A.duranensis, 2n=2x=20, BBand A.ipaënsis, 2n=2x=20, AA) of cultivated peanut and 5S and 45S rDNA as probes, sequential GISH-FISH combined with DAPI staining were employed to develop the karyotypes of Z5163 and the two donors based on distinguishing A and B genome of A. hypogaea clearly. The new karyotype was then used to analyze more peanut cultivars in order to reveal the characteristics of peanut chromosome constitution.【Result】 GISH indicated that all 20 B genome chromosomes of A. hypogaea produced clear and stable signals after hybridized with the total genomic DNA of A.ipaënsis, while only 18 chromosomes except “A chromosomes” of A genome produced signals using A. duranensis as a probe. Sequential mc-FISH using 5S rDNA and 45S rDNA as probes and combined with DAPI staining revealed that all the signals distributed on A and B genome of A. hypogaea were almost identical to its respective possible donor genome chromosomes of A. duranensis and A.ipaënsis, which suggested that A. duranensis and A.ipaënsis were the donors of A. hypogaea. Furthermore, the present study also found that 14 B genome chromosomes showed centromeric bands after DAPI staining, which were quite more than the previous reports, indicating that it was difficult to distinguish the A and B genome chromosomes of A. hypogaea only by DAPI staining as previous reports. Therefore, based on the above findings, a new GISH-FISH karyotype of the cultivated peanut was developed which could clearly distinguish all the chromosomes of A genome from those of B genome in A. hypogaea. And then the karyotype was used to characterize three peanut varieties, and a spontaneous monosomic substitution line of A. hypogaea, MSB1(A1), was found, indicating the homoeologous relationship between chromosomes B1 and A1.【Conclusion】The two genomes of cultivated peanut were very separately correspondent to its possible donor parents A. duranensis and A. ipaënsis; The new karyotype of the cultivated peanut could not only distinguish most of A and B genome chromosomes but also identify spontaneous chromosome variations produced in the progress of human-selection and polyploidization of peanut, indicating homoeologous relationships between chromosomes of A and B genomes of peanut.

Key words: peanut, sequential GISH-FISH, chromosome structure, karyotype, chromosome substitution

DU Pei, LIU Hua, LI Li-na, QIN Li, ZHANG Zhong-xin, HUANG Bing-yan, DONG Wen-zhao, TANG Feng-shou, QI Zeng-jun, ZHANG Xin-you. Chromosome Analysis of Peanut (Arachis hypogaea L.) Based on Sequential GISH-FISH[J].Scientia Agricultura Sinica, 2015, 48(9): 1854-1863.

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Chromosome Analysis of Peanut (Arachis hypogaea L.) Based on Sequential GISH-FISH

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