Kumari Preetesh, Singh Kaushal Pratap, Kumar Sundip, Yadava Devendra Kumar
Indian Council of Agricultural Research (ICAR)-National Institute for Plant Biotechnology, Indian Agriculture Research Institute, New Delhi, India.
Indian Council of Agricultural Research (ICAR)-Directorate of Rapeseed Mustard Research, Bharatpur, India.
Front Plant Sci. 2020 Nov 24;11:575591. doi: 10.3389/fpls.2020.575591. eCollection 2020.
The Brassica coenospeceis have treasure troves of genes that could be beneficial if introgressed into cultivated Brassicas to combat the current conditions of climate change. Introducing genetic variability through plant speciation with polyploidization is well documented, where ploidy augmentation of inter-generic allohexaploids using somatic hybridization has significantly contributed to genetic base broadening. is a member of the Brassicaceae family that possesses valuable genes, including genes conferring resistance to , , pod shattering, heat, and drought stress. This work aimed to synthesize stable allohexaploid (AABBSS) Brassica while incorporating the yellow-seed trait and resistance to stem rot. The two fertile and stable allohexaploids were developed by polyethylene glycol mediated protoplast fusions between (AABB) and (SS) and named as JS1 and JS2. These symmetric hybrids (2 = 60) were validated using morphological and molecular cytology techniques and were found to be stable over consecutive generations. The complete chromosome constitution of the three genomes was determined through genomic hybridization of mitotic cells probed with genomic DNA labeled with fluorescein isothiocyanate. These two allohexaploids showed 24 hybridization signals demonstrating the presence of complete diploid chromosomes from and 36 chromosomes from . The meiotic pollen mother cell showed 30 bivalent sets of all the 60 chromosomes and none of univalent or trivalent observed during meiosis. Moreover, the backcross progeny 1 plant revealed 12 hybridization signals out of a total of 48 chromosome counts. Proper pairing and separation were recorded at the meiotic metaphase and anaphase, which proved the stability of the allohexaploid and their backcross progeny. When screening, the allohexaploid (JS2) of and displayed a high degree of resistance to rot along with a half-yellow and half-brown (mosaic) seed coat color, while the and allohexaplopid1 (JS1) displayed a yellow seed coat color with the same degree of resistance to rot.
甘蓝型油菜拥有大量基因宝库,如果能将这些基因渗入栽培甘蓝型油菜中,对于应对当前气候变化的情况可能会有益处。通过多倍体植物物种形成引入遗传变异性已有充分记载,其中利用体细胞杂交对属间异源六倍体进行倍性增加对拓宽遗传基础有显著贡献。甘蓝型油菜是十字花科的一员,拥有宝贵的基因,包括赋予对菌核病、黑胫病、裂荚、高温和干旱胁迫抗性的基因。这项工作旨在合成稳定的异源六倍体(AABBSS)甘蓝型油菜,同时融入黄籽性状和对菌核病的抗性。通过聚乙二醇介导的甘蓝型油菜(AABB)和诸葛菜(SS)原生质体融合,培育出了两个可育且稳定的异源六倍体,分别命名为JS1和JS2。这些对称杂种(2n = 60)使用形态学和分子细胞学技术进行了验证,发现其在连续几代中都很稳定。通过用异硫氰酸荧光素标记的诸葛菜基因组DNA对有丝分裂细胞进行基因组原位杂交,确定了三个基因组的完整染色体组成。这两个异源六倍体显示出24个杂交信号,表明存在来自甘蓝型油菜的完整二倍体染色体和来自诸葛菜的36条染色体。减数分裂花粉母细胞显示所有60条染色体中有30个二价体,在减数分裂过程中未观察到单价体或三价体。此外,回交后代1植株在总共48条染色体计数中显示出12个杂交信号。在减数分裂中期和后期记录到了正常的配对和分离,这证明了异源六倍体及其回交后代的稳定性。在筛选时,甘蓝型油菜和诸葛菜的异源六倍体(JS)对菌核病表现出高度抗性,种皮颜色为半黄半褐(镶嵌),而甘蓝型油菜和诸葛菜异源六倍体1(JS1)种皮颜色为黄色,对菌核病的抗性程度相同。