| Radish and non-heading Chinese cabbage were important two kinds of vegetables.In this study,radish 'xiaodinghong' was chose as material,induced by different concentration of colchicine,and identified autotetraploid.Finally,we obtained new germplasm of radish.Created a method for early screening in seeding stage to increase doubling rates.The diploid and autotetraploid non-heading Chinese cabbage transcriptional analysis using RNA-Seq was performed.Compared the photosynthetic pathways related genes,and explained why autotetraploid plant photosynthetic capacity stronger than diploid in molecular level.For the future of autotetraploid non-heading Chinese cabbage research provided a data basis.Of colchicine filtered induction autotetraploid seedling traits,increase the rate of doubling,provide the market with high quality autotetraploid radish germplasm.The diploid radish(Raphanus sativus L.)"Xiaodinghong" was induced by treating the apical portion of cotyledon with tconcentrations of colchicine solution by 1.5 g·L-1?2.0 g·L-1 and 2.5 g·L-1,drip 4 and 6 time,respectively.The morphological classification of leaves at two true leaves stage.Using morphology and anatomy and flow cytometry were used to obtain the autotetraploid radish.Agronomic traits,fleshy root nutrition ingredient and germination-related contents of diploid and autotetraploid radish seed were determined.The result showed that the highest mutation rate of treatment group was obtained with 2.5 g·L-1 colchicine solution for 4 times,the highest double rate of treatment group was obtained with 2.0 g·L-1 colchicine solution for 4 times,and the frequency was 5.64%.After colchicine treatment,seedling morphologies was distortion.The seeding leaf response was divided into 'Normal character','Clustered character','Virgate character' and 'Scoop character',the highest rate was 'Clustered character'.Until harvest time,stomata,pollen grain,flowers,leaves and root of the autotetraploid plants showed the 'enormous characteristic'.Compared with diploid plants,stomatal density in single field of view and number of seeds in single pod significant reduction.The content of soluble protein,soluble sugar and Vitamin C in atuotetraploid radish fleshy root were increased23.50%,12.24%and 27.36%,respectively.After two generation of breeding,autotetraploid purity was 95%,and production increased 20.21%.For autotetraploid 'xiaodinghong',compared with diploid radish seed,such as content of soluble protein,soluble sugar and dehydrogenase activly efficiency increased,respectively.Choosing 'Clustered character'could improve the double rate,after continuous progeny selection,we obtained high-quality new autotetraploid radish germplasm.The diploid 'aijiaohuang' and its autotetraploid as material which used Illumina HiSeqTM 2000 platform for transcriptome sequencing.We obtained 70,576 unigenes toally,the average length was 1,108 nt,N50 was 1,626 nt.A total of 66,883 unigenes were annotated.Among them,there were 24,919,56,033 and 37,647 unigenes belonged to COG,GO and KEGG,respectively.The different expression genes(DEGs)in transcriptome were analysized,we obtained 1,769 DEGs.Within them,772 DEGs were up-regulated and 997 DEGs were down-regulated.The KEGG pathway analysis shown that most of DEGs were belonged to metabolic pathways,explained the difference between metabolic level of diploid and autotetraploid.Through to the diploid and autotetraploid non-heading Chinese cabbage transcriptome analysis,provided substantial resources for the study of diploid and autotetraploid,also,provided data for polyploidy genome evolution.This allowed investigation into the divergence of photosynthesis pathway related genes in the non-heading Chinese cabbage diploid and its autotetraploid,to explain the cause of the strengthen photosynthetic ability in autotetraploid.The result show that soluble sugar levels,chlorophyll content,and net photosynthesis rate of the autotetraploid were higher than observed in the diploid,by 29.18%,18.67%,and 19.16%,respectively.1,769 differentially expressed genes(DEGs)were identified.The four photosynthetic regulation pathways from KEGG were analyzed,namely "photosynthesis-antenna proteins","photosynthesis","porphyrin and chlorophyll metabolism",and "carotenoid biosynthesis".KEGG pathway annotation and MapMan analysis indicated that the autotetraploid had a greater ability to absorb light and had a more efficient electron transfer process.Our results indicate that one reason for increased production in the tetraploid was the greater photosynthetic capacity of the autotetraploid.The photosynthetic discrepancy of physiological and molecular data from diploid and tetraploid provides a more comprehensive understanding of the molecular basis of autotetraploid enhanced photosynthetic ability. |
PDF Full Text Request