Research On Physiological And Biochemical And Anther Cell Morphology Of Three Sterile Source Transfected Wucai

Wucai evolved from Braassica,belonging to a variety of the Chinese cabbage.It was cultured widely in the open because of its strong cold resistance,in around the Yangtze River and Huaihe River when it is autumn and winter season.Through after frost,the taste became very crisp and quality was better,so there was a saying ‘wucai is more better than mutton in severe winter' and it was referred to vitamin dish because of containing rich vitamin C.In view of the above-mentioned facts,breeders had done a lot of researches on wucai,but mainly focused on the conventional types of research,such as physiological and biochemical analysis,cultivation techniques and so on.Wucai was cross-pollinated crops,heterosis was very obvious,but breeding technology research started relatively late and lacked of material screening and initiative in terms of disease resistance.All of these factors had limited the breeding new varieties of wucai.This experiment was considered fully the heterosis of wucai and combined with previous researches,studying the agronomic traits and quality trait of F1,which was used with hau type,pol type and wucai cytoplasmic male sterility source,the purpose was to know which kind of infertility source was better;it also analyzed the sterile lines and maintainer line of wucai in the department of flower organ morphology;observating the process of anther development through cytological method and understanding the time and the method of anthers abortive;at the same time we extracted the genome of sterile line and maintainer line to identify the type of cytoplasmic male sterility through(CMS)PCR amplification techniques,hoping to be able to broaden the type of CMS in Chinese cabbage vegetables and promote production and utilization of heterosis in wucai.The study results indicated that :1.Looking from agronomy traits,the offspring of hau type cytoplasmic cabbage infertility source had significant difference in plant height,petiole length,petiole thick,leaf width and weight per plant;the offspring of pol type cytoplasmic cabbage sterile source had no difference in plant width and weight per plant;the offspring of wucai cytoplasmic cabbage sterile source had significant difference in plant height,plant width,petiole length,leaf length,leaf width and weight per plan had significant differences.Compared with backcross parent,the offspring of hau type cytoplasmi cabbage infertility source,expect leaf thickness and leaf width,other characters all had significant difference;the offspring of pol type cytoplasmic cabbage sterile sourcehad significant difference in plant height,plant width,leaf number per plant and leaf width and the offspring of wucai cytoplasmic cabbage sterile source had significant difference in petiole length and leaf width.2.From the point of view of genetic tendency of soluble sugar,soluble protein and vitamin C,soluble sugar was the most stable genetic characteristics no matter what kind of infertility source;soluble protein of wucai infertility source had significant difference during the three period;hau type and pol type genetic trendency were relatively close,there was no significant difference in the first two times.But the soluble protein reached the peak in three infertility source when it was 90 days;the content of vitamin C reached the lowest in 60 days,but no matter what kind of backcross parent of wucai;the margin of change between the maximum and minimum values was 30 times in wucai infertility source,while the range of other infertility source was very large.3.The indexes of floral organ were measured to analyse variability,it can find that there were significant differences between sterile line and maintainer line.And compared with maintainer line 12-14 B,the data of sterile line 12-14 A were significantly smaller.There was a small amount of irregular characters or dyeing light pollen in sterile line's anthers by microscopic observation.Initially,we determined its sterility is relatively stable,and the degree of pollen steritity and rate of sterility were 100% after identification of sterility.4.The anther development of sterile lines 12-14 A was blocked in pollen mother cell meiosis and tetrad stage by observation of paraffin section.Firstly,the pollen mother cells can not develop as normal because of enlargement of tapetal cell,leading to anther abortive.Secondly,the tetrad cells could not split out the normal microspore,microspore appeared sticky united each other and with vacuolization,eventually leading to anther abortive.5.From the stigma of the result of scanning electron microscopy,we could find that the anther of sterile line 12-14 A was shriveled and wizened,and the surface of anther was scale-like,the shriveled and wizened pollen grains were confused together in chamber,the mesh surface of a single pollen grain was damaged,mastoid cells of mature stigma were finger shaped,the surface of these cells was smooth and very full;the anther of maintainer line 12-14 B was rich and its surface was very clear,and the aperture was normal,mesh sizes were different,the surface of net ridge structural was smooth and continuous,the mastoid cell of mature stigma was spoon-shaped.6.The genomic DNA was extracted from sterile line and maintainer line,the length of the strip of gene orf138 was about 460 bp in sterile lines petals amplification products,and had not been found in other organizations.