Prunus Tenella PetSFB6 Gene Cloning And Bioinformatics Analysis And The Population Establishment Of The Regeneration System

Prunus tenella, Rosaceae, Pyunoideae, Amygdalus, It is a unique precious Xinjiang protected species, is a precious ancient Mediterranean Miocene relict deciduous forest trees, have great edible, medicinal and other economic value, and that the gametophytic self-incompatibility affinity of Prunus tenella, affects it's yield and the diffic?Lty of domestication. Prunus tenella have five different populations, and the distribution area of Prunus tenella is diminishing with the increasing of human activities year by year. In this study, the systematic study of five populations of Prunus tenella shoot apex regeneration and regeneration leaf regeneration system of research, this paper for the first time on leaf explants of Five Populations of wild almond callus induction of differentiation test for the future of Xinjiang Prunus tenella Cultivation varieties, lays the foundation of the later work of genetic research and a large number of rapid propagation of seedlings toxic basis, provide a realistic basis for the preservation of high-quality species of seedings. In addition, in this study, the anther of Prunus tenella as material, using RT-PCR and RACE technology to clone PetSFB6 gene, in order to obtain the self incompatible plants provide reliable gene. The main results are as follows:(1)The clone of PetSFB6 gene of Prunus tenellaPrunus tenella cDNA as PCR template PCR, gained about 1300 bp gene, sequencing of the gene for 1223 bp, the largest open reading frame of 1110 bp encoding a protein of 370 amino acids. The gene at Blast than NCBI, and SFB33(HQ148064.3) gene. The similarity of up to 98%. And using the NCBI domain that has the F-box structure, the SFB family genes.(2) Prunus tenella PetSFB6 bioinformatics analysisResults show that the genetic encoding 320 amino acids, speculated that the formulas for the protein C1706H2573N429O470S17, relative molecular mass is 37157.8, the isoelectric point is 6.07, the half-life of theoretical derivation is 0.8 h, negative and positive charge of the total number of amino acid residues respectively 38 and 33, fat index of 83.11, instability parameter is 45.18, the hydrophobic index is 0.145,it is the unstable protein hydrophobicity.(3) The establishment of five populations of Prunus tenella Shoot Tip Regeneration SystemFive different populations of proliferation and the optimum culture medium respectively is: Habahe MS+6-BA 1.5mg/L+IBA 0.1mg/L+GA3 0.5mg/L; Buerjin MS+6-BA 1.0mg/L+NAA 0.1mg/L+GA3 0.5mg/L; Toli MS+6-BA 0.8mg/L+IBA 0.2mg/L+GA3 1.0mg/L; Yu Min MS+6-BA 0.6mg/L L+IBA 0.2mg/L+ GA3 1.0mg/L; Tacheng MS+6-BA 1.0mg/L+IBA 0.1mg/L+GA3 0.5mg/L;. Among them, five populations of stem apex proliferation of the optimal sucrose concentration is different, Buerjin, Tacheng, Habahe, the best concentration of sucrose was 30g/L Yu Min, Torli 35g/L.The five group in rooting shoots the optimal hormone combination for the Habahe 1/2MS+6-BA 0.04mg/L+IBA 1.5mg/L. Buerjin 1/2MS+ 6-BA 0.04 mg/L + IBA 0.6mg/L; Toli 1/2MS+ 6-BA 0.04 mg/L + IBA 0.8mg/L; Yu Min 1/2MS+ 6-BA0.04 mg/L + IBA 1.0mg/L; Tacheng 1/2MS+ 6-BA0.04 mg/L + IBA 1.2mg/L. Due to the different habitats of the five populations, the best rooting hormones also differences.(4) The establishment of five populations of Prunus tenella leaf regeneration system.Five in the induction group Prunus tenella leaf callus, callus induction of the best Buerjin leaves dark culture time was 21 d, petiole, middle leaf, and leaf apex the ability of callus induction from big to small order: petiole >middle leaf > leaf apex; Five populations of Prunus tenella leaves the best callus induction medium was: Buerjin NN69+ZT 2 mg/L+IBA 0.3 mg/L; Habahe MS + ZT 2mg/L + IBA 0.2mg/L; Yu Min MS + ZT 3mg/L + IBA 0.2 mg/L; Toli MS + ZT 4mg/L + IBA 0.3mg/L; Tacheng MS + ZT 4mg/L + IBA0.2mg/L.Buerjin in the induced group Prunus tenella leaf buds, when cultured in cytokinin / auxin ratio is high, the increase of ZT concentration in Buerjin leaf bud induction rate showed a parabola trend, cytokinin plays a leading role, to induce buds differentiation, cytokinin(ZT) inhibit the bud differentiation concentration is highly, the best medium of leaf bud in Buerjin induction is: NN69 + ZT 4 mg/L + IBA 0.05mg/L.