| Peanut,also known as groundnut,is an important oil and cash crop that is planted worldwide.Peanuts are versatile and play an important role in our lives.However,the yield and quality of peanut are affected by drought,and drought is the main limiting factor of peanut production in China.AREB1(Abscisic-acid respons element binding protein 1)is a class of drought-related transcription factors.This research group has cloned and analyzed function of AhAREB1 gene from peanut.In this study,another gene of AhAREB1 family was cloned from feng hua 2.The relationship between expression quantity of peanut AhAREB1-2 gene and drought resistance was studied under the condition of osmotic stress.At the same time,the silent vector of AhAREB1-2 was constructed,and the agrobacterium-mediated pathway was transferred to feng hua 2 to obtain the silent gene plant.The main findings are as follows:1.Cloned the two CDS sequences of the gene Ah AREB1-2 from feng hua 2 by RT-PCR,named AhAREB1-2a and AhAREB1-2b gene.They encode 370 and 421 amino acids.The software predicted that the molecular weights of the two proteins were 38.7kda and 44.9kda respectively,and the isoelectric points were 9.90 and 9.47 respectively.Conservative structural analysis shows that AhAREB1-2a and AhAREB1-2b protein have a conservative structural domain of C1,C2,C3,C4 and bZIP.2.The tissue specificity of AhAREB1-2a and AhAREB1-2b gene was analyzed and the induced expression pattern was observed in 5 abiotic stress.Results of tissue-specific showed AhAREB1-2 gene were expressed in shoots,roots,leaves and seeds,but the highest expression level in the leaves.Peanut seedlings were treated by five kinds of abiotic stress,we found that AhAREB1-2 gene was induced by ABA(100μM),dehydration drought,high salt(250mmol/L NaCl),15% PEG6000 and MV(50μmol/L),and 15%PEG6000 has the highest expression leaves at 4 hours.Analysis of 26 varieties(or lines)in seedling leaves AhAREB1-2 gene expression under 15% PEG6000 stress after 4 hours,the results showed that the drought resistance coefficient of peanut varieties was positively correlated with the expression of gene AhAREB1-2.3.Using the method of double enzyme cutting and connection,the plant silent vector TRV2-AhAREB1-2a and AhAREB1-2b,which contain AhAREB1-2a and AhAREB1-2b gene,were constructed with TRV2 as the basic vector.The gene silencing plant was obtained through the genetic transformation of peanut by agrobacterium GV3101.The gene silencing plants and in vitro leaf was treated with 15%PEG,250mmol/LNaCl,and 50 mol/L MV stress.Observed phenotypic appearance change the results show that the natural drought and PEG6000 stress treatment silence after processing plant wilting degree is higher than the control plant,loss of green velocity of vitro leaf faster than control plant;The rate of loss of green in the vitro leaves of the silent plant was higher than that of the control plant after high salt stress.The degree of oxidation in the vitro leaves of the silent plant was more serious than that of the control plant after MV stress.The water loss rate of the plant leaves in vitro was higher than that of the control.The water loss rate is greater in the vitro leaves of the silent plant than that of the control plant.It is indicated that the AhAREB1-2a and AhAREB1-2b gene play an important role in the drought resistance,salt tolerance and antioxidant pathway of peanut. |
PDF Full Text Request