Analysis Study Of Hot Pepper (Capsicum Spp.) With Botanical Character And RAPD

Twenty eight hot pepper materials were adopted and their botanical traits for example the geniculum of protanthesis,the average height, the length of fruit, the width of fruit, the thickness of sarcocarp and the weight of fruit were surveyed. CTAB was applied to extraction material DNA. The RAPD condition was optimized. Then twenty eight hot pepper materials were clustered with botanical traits and RAPD. The results follows:1. The improved CTAB was applied to extract DNA. The ratio of OD260/OD280 was from 1.8 to 2.0, and OD260/OD230 was more than 2. All above indicated that the DNA was qualified for RAPD analysis.2. Single factor of RAPD was optimized one by one, then set up optimized RAPD reaction. Each sample was amplified in a reaction mixture (20μL in total volume) containing 1.0U of Taq polymerase, 0.2mmol/L of dNTP, 2.0mmol/L of MgCl₂, 0.2μmol/L of random primer, 80ng template DNA, 2μl of 10×PCR buffer, and the other was ddH₂O. The PCR tip with the reaction mixture was covered by 10μl of mineral oil. The reaction mixture was cycled through the following temperature profiles: 94℃for 4min, 94℃for 1min, 37℃for 1min, and 72℃for 1.5min for 40 cycles. The PCR was progradated at 72℃for 7min, and stored at 4℃.3. The 13 primers were screened out of 38 primers. The 13 primers amplified 157 DNA fragments and each primer could amplify 12.1 fragments. There were 128 polymorphic fragments among total 157 fragments, which accounted for 81.52% and each primer could amplify 9.9 polymorphic fragments average.4. Cluster results of the morphology of 28 varities were divided into five categories. 13 materials (NO.20-Chaotianjiao- 4 , NO.21-Chaotianjiao- 5, NO.23-Chaotianjiao- 7 , NO.2-Yeshanjiao-1,NO.22-Chaotianjiao-6,N0.26-ShanDongqixing and NO.3 -Yeshanjiao-2,NO.4-Zisewucaijiao,NO.17-Chaotianjiao-l,NO.19-Chaotianjiao-3,NO.24-C haotianjiaotianmi,NO.18-Chaotianjiao-2 and NO.8-Erjintiao) belonged to group one; 7 materials (N0.7-Huixiang, N0.25-Hongcaijiao, NO.9-Changjiao-7^#, NO.12-Lajiao-118^#, NO.27-Kangbing-1^#, NO.13-Xinhong and NO.28-Jiuxiang) belonged to group two; 6 materials (NO.10-Changjiao-2-1, NO.14-Huangyu, NO.16-Shengli and NO.15- Dongfang NO.5-Changlajiao and NO.6-Qianhui) belonged to group three; NO.1-Mosxige belonged to group four, NO.11-Luojiao-98^# belonged to five.5. Analysised by NTSYSpc, 28 pepper materials were divided into six major categories in similar coefficient of 0.780. NO.1 belonged to group one; 10 materials(NO.16-Shengli, NO.7-Huixiang, NO.13-Xinhong, NO.14-Huangyu, NO.6-Qianhui, NO.5-Changlajiao, NO.8-Erjintiao, NO.9-Changjiao-7^#, NO.12- Lajiao-118^# and NO.10- Changjiao-2-1) belonged to group two; NO.11 belonged to group three, 4 materials(NO.2-Yeshanjiao-1, NO.4- Zisewucaijiao, NO.3- Yeshanjiao -2, NO.15-Dongfang) belonged to group four, 11 materials(NO.26- ShanDongqixing, NO.22-Chaotianjiao-6,NO.21-Chaotianjiao-5,NO.24-Chaotianjiaotianmi,NO.25- Hongcaijiao, NO.20- Chaotianjiao-4, NO.23- Chaotianjiao-7, NO.17- Chaotianjiao-1, NO.19-Chaotianjiao-3, NO.18-Chaotianjiao-2, NO.28-Jiuxiang) belonged to group five, NO.27 belonged to group six.