Identification of S-RNase genotype and analysis of its origin and evolutionary patterns in Malus plants

Identification of the S genotype of Malus plants will greatly promote the discovery of new genes,the cultivation and production of apple,the breeding of new varieties,and the origin and evolution of self-incompatibility in Malus plants.In this experiment,88 Malus germplasm resources,such as Aihuahong,Xishuhaitang,and Reguanzi,were used as materials.Seven gene-specific primer combinations were used in the genotype identification.PCR amplification using leaf DNA produced a single S-RNase gene fragment in all materials.The results revealed that 70 of the identified materials obtained a complete S-RNase genotype,while only one S-RNase gene was found in 18 of them.Through homology comparison and analysis,13 S-RNase genotypes were obtained:S_(1)S_(2)(Aihuahong,etc.),S_(1)S_(28)(Xixian Haitang,etc.),S_(1)S_(51)(Hebei Pingdinghaitang),S_(1)S_(3)(Xiangyangcun Daguo,etc.),S_(2)S_(3)(Zhaiyehaitang,etc.),S_(3)S_(51)(Xishan 1),S_(3)S_(28)(Huangselihaerde,etc.),S_(2)S_(28)(Honghaitang,etc.),S_(4)S_(28)(Bo 11),S_(7)S_(28)(Jiuquan Shaguo),S_(10)S_e(Dongchengguan 13),S_(10)S_(21)(Dongxiangjiao)and S_(3)S_(51)(Xiongyue Haitang).Simultaneously,the frequency of the S gene in the tested materials was analyzed.The findings revealed that different S genes had varying frequencies in Malus resources,as well as varying frequencies between intraspecific and interspecific.S_(3) had the highest frequency of 68.18%,followed by S_(1)(42.04%).In addition,the phylogenetic tree and origin evolution analysis revealed that the S gene differentiation was completed prior to the formation of various apple species,that cultivated species also evolved new S genes,and that the S_(50) gene is the oldest S allele in Malus plants.The S_(1),S_(29),and S_(33) genes in apple-cultivated species,on the other hand,may have originated in M.sieversii,M.hupehensis,and M.kansuensis,respectively.In addition to M.sieversii,M.kansuensis and M.sikkimensis may have also played a role in the origin and evolution of some Chinese apples.

Nitric Oxide Regulates Mitochondrial Fatty Acids and Promotes CBF Expression of Peach Fruit during Cold Storage

Low-temperature storage is convenient for postharvest preservation of peach fruit,but peach fruit is sensitive to cold damage,which lowers its quality.Nitric oxide(NO)has the potential to improve the bitter resistance of peach fruit.In this work,peach fruit was treated with 15μmol L^(−1)NO and 5μmol L^(−1)c-PTIO[2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxo-3-oxide],to study changes in mitochondrial fatty acids and expression of the C-repeat binding factor(CBF).The results showed that 15μmol L^(−1)exogenous NO significantly maintained fruit quality,reduced peroxidation of mitochondrial fatty acids,increased the activities of the antioxidants superoxide dismutase(SOD),peroxidase(POD),catalase(CAT),ascorbic acid peroxidase(APX),and reduced the content of hydrogen peroxide(H_(2)O_(2)).Meanwhile,NO treatment suppressed the increase in browning index and ion leakage rate,increased the activity of S-nitrosoglutathione reductase(GSNOR),the contents of S-nitrosothiols(SNOs),and the ratios of mitochondrial NAD^(+)/NADH and NADP^(+)/NADPH,increased the expression levels of PpCBF1/5/6.However,the expression levels of PpCBF2/3/4 were not significantly regulated by exogenous NO.Peaches treated with c-PTIO showed opposite effects to those treated with exogenous NO.These results suggest that exogenous NO can improve antioxidant capacity,preserve mitochondrial fatty acids,and upregulate the expression of PpCBF1/5/6 to alleviate cold tolerance and maintain the peach quality during storage.