The artificial allotetraploid of Gossypium herbaceun and G.raimondii was created in the laboratory to broaden the ge-netic background of cotton and provide novel materials for the germplasm innovation of cotton.In this study,fiber characters,physiological and biochemical comparison,SSR markers and floral transcriptomes analysis were conducted among three genera-tions of the artificial allotetraploid and their parents,including G hirsutum L.acc.TM-1.The results showed that the fiber color of artificial allotetraploid,light yellow,mid-parent of G herbaceun and G.raimondii,and the fiber length was close to TM-1.The activities of SOD,POD,CAT,and MDA content in artificial allotetraploid plants were significantly higher than parents and TM-1,indicating that the stress resistance of artificial allotetraploid was enhanced after distant hybridization and chromosome doubling.55 polymorphic loci were detected from 11 pairs of SSR primers,with 2-10 mutation loci per primer and an average of 5.PIC ranged from 0.498 to 0.892,with an average of 0.742.The results indicated that SSR markers could reflect rich genetic diversity information among the test materials.The complementary of parents and specific bands were amplified in S2,S3,and S4,simulta-neously,indicating that chromosome segment recombination occurred in the artificial allotetraploid at the molecular level.There were 37,19,and 20 bands of S2,S3,and S4 identical with TM-1 respectively,among which 17 bands were shared with TM-1 by all generations,indicating that the artificial allotetraploid had homology in genetic background with TM-1.The floral organ tran-scriptome analysis showed that there were 5653 differentially expressed genes(DEGs)between artificial allotetraploid and TM-1,in which 3062 were up-regulated and 2591 were down-regulated.GO functional analysis showed that DEGs had significant differences in photosynthesis,photosystem Ⅱ,RNA directed DNA polymerase activity and other pathways.KEGG enrichment analysis showed that DEGs were more active in photosynthetic-antenna proteins,photosynthesis and isoflavone biosynthesis.The enrichment analysis of fertility related DEGs between artificial allotetraploids and TM-1 revealed that most of the DEGs of artificial allotetraploids in pollen development and recognition were down-regulated,which may be the key factor leading to low seed setting percentage of artificial allotetraploid.Three G-type lectin receptor serine/threonine protein kinases(Gh_D01G067500,Gh_D05G168100,Gh_A01G062500)were obtained from Nr annotation information by clustering.The results showed that the artificial allotetraploid performed advantage in fiber color,resistance to stress and photosynthesis,combining the excellent quality of its parents.It is ex-pected to restore its fertility by crossbreeding with G hirsutum,and to further cultivate a valuable novel material for production.
关键词
人工异源四倍体/生理生化测定/SSR分子标记/转录组测序/差异表达基因
Key words
artificial allotetraploid/physiological and biochemical detection/SSR molecular marker/transcriptome sequencing/differentially expressed gene
维普
万方数据