Peanut(Arachis hypogaea L.),an improtant oil crop,usually encounters drought stress in the process of growth and development,especially at pre-flowering stage. In order to gain insight into the drought tolerance potentials based on osmolyte accumulation and metabolism of proline aspects of peanut,pot experiments were conducted with a split-plot design in Tai'an,Shangdong Province,China in 2013 and 2014. Pre-flowering drought(PFD) stress and optinum irrigation(control,CK) were served as the main plots and the two peanut cultivars Shanhua 11 and Hua 17 served as sub-plots. Shanhua 11 was drought-tolerant cultivar and Hua 17 was drought-sensitive. The content of soluble sugars,soluble protein,free proline and other free amino acids,the activities of enzymes involved in proline metabolism,and malondialdehyde(MDA) content and ion leakage were all investigated in the two cultivars at pre-flowering stage. Results showed that PFD stress significantly increased the levels of soluble protein,free proline and free amino acid,and increased Δ1-pyrroline-5-carboxylate synthetase(P-5-CS,EC 2.7.2.11) activity in the leaves of drought-tolerant and drought-sensitive cultivars. The activity of proline dehydrogenase(pro DH)(EC 1.5.99.8) decreased under PFD stress in both cultivars. The leaves of the tolerant cultivar maintained higher increments of osmolyte levels,lower increments of MDA content and ion leakage,as well as a higher increased proportion of P-5-CS activity and higher inhibited proportion of pro DH activity under water stress compared with the drought-sensitive cultivar. The study suggests that proline accumulation in peanut leaves under PFD can be explained by the higher enhanced activities of P-5-CS and higher inhibition of pro DH. The results will provide useful information for genetic improvement of peanut under drought tolerance.
Superoxide dismutase(SOD, EC 1.15.1.1) plays a key role in response to drought stress, and differences in SOD activity changes among cultivars are important under drought conditions. We obtained the full-length DNA of the chloroplast Cu/Zn-SOD gene(Ah CSD2)from 11 allotetraploid cultivars and 5 diploid wild species in peanut. BLAST search against the peanut genome showed that the Ah CSD2 genes g CSD2-1 and g CSD2-2 are located at the tops of chromosome A03(A genome) and B03(B genome), respectively, and both contain 8exons and 7 introns. Nucleotide sequence analyses indicated that g CSD2-2 sequences were identical among all the tested cultivars, while g CSD2-1 sequences showed allelic variations.The amino acid sequences deduced from g CSD2-1 and g CSD2-2 both contain a chloroplast transit peptide and are distinguished by 6 amino acid(aa) residue differences. The other 2aa residue variations in the mature peptide regions give rise to three-dimensional structure changes of the protein deduced from the genes g CSD2-1 and g CSD2-2. Sequences analyses of cultivars and wild species showed that g CSD2-2 of Arachis hypogaea and g Aip CSD2(Arachis ipaensis) are identical, and despite the abundant polymorphic loci between g CSD2-1 of A.hypogaea and sequences from A genome wild species, the deduced amino acid sequence of Ah CSD2-1(A. hypogaea) is identical to that of Adu CSD2(Arachis duranensis), whereas Aco CSD2(Arachis correntina) and Aca CSD2(Arachis cardenasii) both have 2 aa differences in the transit peptide region compared with Ah CSD2-1(A. hypogaea). Based on the Peanut Genome Project, promoter prediction revealed many stress-related cis-acting elements within the potential promoter regions(pp-A and pp-B). pp-A contains more binding sites for drought-associated transcriptional factors than pp-B. We hypothesize that the marked changes in SOD activity in different cultivars under drought stress are tightly regulated by transcription factors through transcription and expression of Ah CSD2 genes.
Xiurong ZhangQian WanFengzhen LiuKun ZhangAiqing SunBing LuoLi SunYongshan Wan
