Single-pollen genotyping to estimate mode of unreduced pollen formation in Citrus tamurana cv. Nishiuchi Konatsu
Single-pollen genotyping to estimate mode of unreduced pollen formation in Citrus tamurana cv. Nishiuchi Konatsu
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DOI：
10.1007/s00497-016-0277-7
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发表时间：
2016-03
影响因子：
3.4
通讯作者：
C. Honsho;A. Sakata;Hikaru Tanaka;S. Ishimura;T. Tetsumura
中科院分区：
文献类型：
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作者：
C. Honsho;A. Sakata;Hikaru Tanaka;S. Ishimura;T. Tetsumura
柑橘FDR花粉形成的关键信息。来自Tanaka; NK），具有产生未减数的2n花粉粒的能力，允许通过有性多倍体化产生多倍体后代。本研究建立了柑橘单花粉基因分型方法，并将其应用于NK 2n花粉杂合性传递的分析。杂合性传递表示为从亲本到2n配子的一组杂合等位基因的遗传百分比。通过将观察到的杂合性传递和遗传距离应用于第一次分裂恢复（FDR）和第二次分裂恢复（SDR）两种不同的图函数，研究了2n花粉发育的途径。计算两个函数的观测值的拟合，同时虚拟地移动着丝粒位置。我们在NK中筛选了6个杂合SSR（共显性微卫星标记位点），所有这些位点都位于同一连锁群内。花粉萌发前的DNA提取是这项工作所必需的，和6小时孵育被证明是最佳的后续PCR扩增。单花粉基因分型未减少NK 2n花粉粒显示，杂合性传输超过50%，在所有6个等位基因，和健身测试表明，FDR地图功能更好地拟合杂合性传输观察到的，而不是SDR功能。因此，我们的数据强烈表明，2n花粉在NK是第一次分裂恢复的结果。
Key message2n pollen formed by FDR in citrus.AbstractThe Japanese local citrus cultivar, Nishiuchi Konatsu (Citrus tamuranahort. ex Tanaka; NK hereafter), has the ability to produce unreduced 2n pollen grains, allowing generation of polyploid progenies via sexual polyploidization. In this study, we developed a method of single-pollen genotyping for citrus and applied it to the analysis of transmission of heterozygosity in NK 2n pollen grains. Heterozygosity transmission was expressed as the percentage inheritance of a set of heterozygous alleles from the parent to the 2n gamete. The pathway of 2n pollen development was investigated by applying the observed heterozygosity transmission and genetic distance to two different map functions, for first division restitution (FDR) and second division restitution (SDR). The fit of the values observed for both functions was calculated, while virtually moving the centromere position. We screened for six heterozygous SSR (codominant microsatellite marker loci) in NK, all of which were expected to lie within the same linkage group. Pollen germination prior to DNA extraction was essential for this work, and 6-h incubation proved to be optimal for subsequent PCR amplification. Single-pollen genotyping unreduced NK 2n pollen grains revealed that heterozygosity transmission exceeded 50 % in all six alleles, and fitness tests indicated that the FDR map function better fitted the heterozygosity transmission observed rather than the SDR function. Our data thus strongly indicate that 2n pollen in NK is a result of first division restitution.