Study On Cold Resistance And Molecule Detection Of Progenies Of CBF3 Transgenic Eggplant

Eggplant (Solarium melongena L.) is a thermophilic vegetable,22 to 30 ℃ is the best temperature for its growth. Chilling injurying often happens in winter and spring when temperatures are low, causing growth retardation and reduction both in production and quality. Chilling injurying occurred in 0 to 10 ℃ at a low temperature hinders normal physiological activities of the plant. The temperature and the duration of the low temperature determined the degree of damage to the plants. Anti-season vegetables cultivation is increasing as delaying growth till autumn and planting vegetables in winter and spring bring remarkable economical benefit. An increasing trend of anti-season vegetables cultivation highlights the importance of cold-resistant varieties. However, The varieties of eggplant with good cold resistance are not much. The high quality seed prices from abroad are high, the cold resistance is limited, and the consumption habits are not suitable. Although more researches have been done on cold resistance character of tomato and pepper, less study on eggplant.Arabidopsis CBF1, the activating transcription factors of COR cold (cold regulated), increases an important plant genetic engineering. This kind of transcription factors can be combined with the core components of COR CRT/DRE specificity so as to enhance downstream gene expression and stimulate diverse resistance mechanism of plants, improve the transgenic plants of resistance against low temperature, drought, salinity and other abiotic stress. Based on this, many scholars have conducted researches on gene transformation of Arabidopsis CBF1, and have obtained the ideal materials about resistant transgenosis. All these work provide a theoretical basis for researches on plants of resistance against low temperature, drought, salinity and so on.Based on the having trans-gene AtCBF3 eggplants, this research is conducted on and discussed from the cultivation of transgene T1 eggplants by self-crossing, aiming to investigate whether exogenous gene is expressed in trans genetic plant offspring and what are the differences in biological characteristics, and provide theoretical basis for the breeding of new varieties of trans genetic eggplant.Taking four lines of transgenetic eggplant (B4, B27, A16, A27) and their offspring as well as wild-type control CK (the second generation) as experiment materials, the research is mainly focused on the following aspects:(1) To detect T2’s transgenic line and its descendants on PCR、RT-PCR and explore whether the exogenous gene is steadily inherited in the offspring of transgenic plant;(2) To test physiological and biochemical indexes such as the relative conductivity, the malondialdehyde (MDA), catalase (CAT) and proline of descendants of transgenic plants who tested positive for PCR and wild type control CK by setting them under low temperature;(3) To conduct cold tolerance identification on transgenic plants and their offspring. Then get the following conclusion:1. By detecting T2’s transgenic lines B4、B27、A16、A27 and wild type control CK for PRC testing, the result comes out that gene AtCBFS was successfully switched into genomes of B4 and A27;2. With the treatment under 4℃ low temperature, the plasma membrane stability and related indicators were detected. The relative conductivity and MDA of transgenic plants was lower than those of wild type controls, whereas the CAT transgenic plants which can clear reactive oxygen are higher than that of wild type controls. All the results show that AtCBF3 has played a vital role in the stability of the plasma membrane;3. With the treatment under 4℃ low temperature, the content of osmotic regulation substances in the cell was measured, and found that the transgenic plants were higher than case-control in terms of content of soluble protein, soluble sugar and proline. The results showed that the excess of AtCBF3 gene expression is helpful to maintain the cell’s osmotic pressure;4. With the treatment under 4℃ low temperature, the anti cold ability between transgenic plants and wild type eggplants was detected. The chilling injury degree of transgenic plants was significantly lighter compared with wild type controls, and this clearly shows that the excessive expression of genes is the direct factor of improving cold resistance degree of transgenic eggplants;5 With membership function method to carry on the comprehensive analysis of the physiological and biochemical indexes, transgenic plants A27, B4 and wild type control CK have the order with B4>A27>CK of the cold resistance.