| Cotton is one of the most important economic crop in China. However, Verticillium wilt of cotton, which is one of the most dangerous fascicular disease caused by Verticillium dahliae Kleb., is the major diseases which seriously affect the yield and quality of cotton. Although much progress has been made in prevention and control of Verticillium wilt of cotton, the progess is slow. Because Verticillium dahliae is soil-borned fungal pathogen, the effect of prevention and contrail is very little using the physical and chemical method. It is strongly consided that developing resistant and tolerant cultivar is the most economic and effective ways to resolve this problem. But the basic research about the pathogenic mechanism of Verticillium dahliae shows that developing resistance and tolerant cultivar by conventional breeding is far from enough because lack of resistant resources.A specific polyclonal antiserum was prepared again a purified phytotoxin from Verticillium dahliae of cotton, and an indirect EL1SA procedure was developed for detection of Verticillium dahliae Phytotoxin (PLPC) in culture filtrates. Results showed that the PLPC in culture filtrates was detectable when 11 strains were grown in Czapek's medium under shaking condition at 25 ℃. only for 24 hours, which is 3 days earlier than that of the bioassay. The ability that different character strain produced phytotoxin was different. The average phytotoxin concentration of defoliating strain was lower than that of strain which was not defoliating in the other two days. But from the third day, the ability of that defoliating strain produced phytotoxin was higher than it of strain which was not defoliating. The phytotoxin concentration of five strains (V991, V43.1, T9, V250, V123) were the highest in the ninth day; other five strains (V232, VB, V56, V120, V59) produced phytotoxin the most in the eleventh day; the phytotoxin concentration of VJVII was the highest in the seventh day.Using the susceptive cultivar (Junmian 1) and different concentration phytotoxin of V991 to detect cotton seedling depending on the method of root dipping in phytotoxin, the result was that all of the different concentration phytotoxin of Verticillium dahliae could cause cotton wilt; in a certain degree, the higher concentration phytotoxin could cause cotton wilt more seriously. 1/8 times of phytotoxin was fit for early diagnosis of Verticillium wilt of cotton and it was used to detect the wilt difference among the strains (V991, V56, V232, V59) . As a result, we found that the phytotoxin of Verticillium dahliae could be used to judge the character of cultivar in diagnosis and the wilt difference of Verticillium dahliae in experiment. In a certain period, the longer the experiment time was, the higher the wilt index was. When cotton seedling was treated 36h~48h, the difference of every strain was clear and the wilt index of defoliating strain was higher than that of strain which was not defoliating.Through method of root dipping in phytotoxin, 1/8 times of phytotoxin was used to detect the character of Wenwu, 9456D, X033 (Gossypium hirsutium) , searisland cotton 7124 (G.. barbadense) , Junmian 1 in experiment. It was found that the wilt index of different cultivar would change as the experimental time increasing. When experimental time was 24 h, the wilt index difference of cultivars was 22.77~24.77. When experimental time was 36 h, the difference was 34.53~43.81. But experimental time should be 36~48 h. The experimental result showed that the wilt index of different cultivar was apparently different. Whats'more, it could utlize the phytotoxin of Verticillium dahliae to differentiate the different cultivar.The four methods of total protein extraction in cotton leaves, three methods of SDS-PAGE and three different concentration detaining were compared and improved. The results showed that it could make better to utilize 25 mmol L~-1, pH8.0 Tris-HCl on total protein extraction in cotton leaves. Using this method, the protein was extracted completely and efficiently. Moreover, satisfied results were achieved in SDS-PAGE. The method of modified pyruvic subside in SDS-PAGE in cotton leaves could be used to test toothful and the clear electrophoresis result could be obtained. When it was used modified pyruvic subside in SDS-PAGE, confecting different consistent separate gel, the consisitence of 10 %~12 % fit for SDS-PAGE of total protein in cotton leaves. Through SDS-PAGE, A conclusion was found that the temperature related to destaining: the higher temperature, the shorter time of destaining, within some range temperature. The experimental cultivars were the ones which were resistance cultivars: Wenwu, 9456D, X033 (Gossypium hirsutium) , sea-island cotton 7124 (G. barbadense) and susceptive cultivar: Junmian 1 and which were induced by Verticillium dahilae, another ones which were planted without pathogeny, such as Wenwu, 9456D, X033, sea-island cotton 7124, Junmianl. Depending on SDS-PAGE, it was found there was one more trap in the protein induced by Verticillium...
|
PDF Full Text Request