Identification Of Jute(Corchorus Capsularis L.) Infertility Type And Analysis Of Differences In Protein Expression

Jute which belongs to the Tiliaceae, Corchorus, is a short-day, annual, herbaceous plant cultivated for the soft bast fiber in the stem. It has the advantages of excellent environment-stress tolerance, wide adaptability, high fiber yield and good quality.This research took the Jute 179, Yue Yuan 6 and the sterile mutant as test materials, morphological observation of pollen grain, iodine-dye assay, sprouting of pollen assay in vitro and pollination experiment in field were carried on to indentify the mutants were high temperature sensitivity male-sterility and female-sterile mutant. In bud previous stage and bud period, determination of physiological and biochemical indexes and analysis of proteomics were taken to reveal sterile mechanism and to know the differences between the jute Yue Yuan 6 sterile mutant and its normal materials. This research obtained the results as following:1. The relationship between the pollen germination rate and temperature were researched,the optimum temperature of jute pollen germination was 28—30℃in vitro, in other temperature, the pollen germination rate decreased as the temperature getting up or down, when it getted upper limit temperature, the pollen didn't germinate. At the optimum temperature, the pollen began to germinate after culture 1 hour,and the germination rate was maximum after 3 hours.2. Sprouting of pollen assay in vitro and pollination experiment in field were taken to analyze vitality of pollen of Yue Yuan 6 fertile and infertile mutant materials,which showed the pollen of infertile mutant material had the nomal vitality; It found that the infertile mutant didn't have The normal development of the stigma when anatomized floral organ at the same period of the two materials. So Yue Yuan 6 sterile mutant was female-sterile due to the stigma was premature degradation. 3. The content of the soluble sugar and the soluble protein of Yue Yuan 6 fertile material and the sterile mutant material in bud previous stage were compared, bud phase and productive phase. The results showed that the soluble sugar and the soluble protein displayed the dynamic change differently along with the growth time in functional leaves and flowers between the two materials. The contents of soluble sugar in the sterile mutant materials were lower than the fertile materials in bud previous stage in functional leaves; however, it was opposite in bud phase and productive phase, these two values were 16.0% and 33.3%, respectively. In bud tissue of sterile mutant materials the contents of soluble sugar was lower than the fertile materials, and the value was 15.5%. The soluble protein's expression quantity in functional leaves and bud tissues of the fertile materials were higher compared to the sterile mutant materials in 3 different periods, the values were 3.9%, 29.2%, 79.1%, 11.9%, respectively. The expression of metabolic difference of above-mentioned may be related to the female sterile.4. Protein extraction system for jute leaves have established, TCA-acetone extrac- tion from jute leaves is more efficient than the method of Tris-base acetone and Tris- HCl extraction and the protein spots were significantly increased. And the protein extraction ratio reached 15.8% at most by TCA acetone method, and 44.2% and 114.1% higher than by other two methods, respectively. Besides, the 2-DE image background was the clearest. The effect of the sample preparation pyrolysis in ultrasonic 40 min was much better than in 32℃constant temperature water-bath. The construction of two-dimensional electrophoresis system can be use to study the differences of protein expression on normal material and mutant of jute, and promote the application of proteomics in jute mutant.5 Different proteins between the Yu Yuan 6 fertile material and the sterile mutant material were compared, picked out 22 protein spots to do MALDI-TOF-MS/MS tandem mass spectrometry analysis from 46 discrepancy proteins.of which 15 protein spots can be identified. 7 protein spots were down in sterile mutant and 4 protein spots were up in sterile mutant material, others spots were dysregulated spots. The name of those 15 protein as following: Rubisco(4), nascent polypeptide associated complex alpha subunit, Putative cytochrome c oxidase subunit, ATPase, phosphoglycerate mutase, triosephosphate isomerase communis, NAD dependent epimerase/dehydr- atase, ribosomal protein S14, dihydrolipoamide dehydrogenase, small heat shock protein, Light-induced protein, Carbonic anhydrase. The function of the proteins involved the material energy, sugar metabolism, protein degradation, metabolic processes, the female-sterile of jute is possible related those metabolic system.