Function Analysis Of The Sorbitol-6-phosphate Dehydrogenase Gene Promoter From Apple

Sorbitol. is the primary.photo.synth.etic produ.ct in many spec.ies of Rosaceae. In addition, sorbitol is an efficient osmolyte. Its accumula.tion can impro.ve plant tolera.nce against abiotic stress.NADP-depende.nt sorbit.ol-6-phosph.ate dehydroge.nase(S6PDH)play a key role in the biosynth.esis of sorbitol..In the previ.ous study, we found that the expressi.on of S6 PD.H promo.ter(S6PDHp)may be ti.ssue-specific and stress.-inducible. In this study, Arabidopsis mediated method GUS S6 PDHp and deletions fusion expression vectors were transformed into tobacco, Arabidopsis, tomato. And received positive transgenic plants. S6 PDHp function of a preliminary analysis. The research results proved S6 PDHp functional characteristics and know S6 PDH regulation of gene expression provides a theoretical basis. Main results are as follows:1. In this stu.dy, ‘Cata.lpa child’ apple(Malus pruni.folia Wil.ld.Borkh) for the materi.al, the S6 PD.H clon.ed gene prom.oter seque.nce and its differ.ent lengt.hs of promoter. 5 ’end deletio.ns. They are S6PDHp1-2.396 bp, S6PDHp2-2122 bp, S6PDHp3-1719 bp, S6PDHp4-1066 bp, S6PDHp5-836 bp and S6PDHp6-236 bp. The S6 PDHp deletio.ns and pC0390-GUS was construct.ed using two restricti.on enzymes S6 PDH gene promoter GUS gene transie.nt expressi.on vector plants. By transie.ntly transfor.med tobacco seedlings GUS activity analysis, we found that six cloned promoter deletions are having activity. And to transiently transformed tobacco ABA, MeJA, salicylic acid, 4 ℃ cold, dark and GA3 stress treatment. Under the SA treatment, GUS-S6PDHp-2396, GUS-S6PDHp-2122, in response to GUS-S6PDHp-1719, and GUS-S6PDHp-646 is significantly. In the ABA treatment, the response GUS-S6PDHp-1066 and GUS-S6PDHp-251 is significantly. In the dark treatment, the only response GUS-S6PDHp-2396 significantly. In the process the next GA, GUS-S6PDHp-2396, in response to GUS-S6PDHp-1066 and GUS-S6PDHp-646 is significantly. Under the methyl jasmonate, GUS-S6PDHp-2122, in response to GUS-S6PDHp-1066 and GUS-S6PDHp-251 is significantly.2. Saved use of the laboratory, ‘Gala’ sorbitol-6-phosphate dehydrogenase gene promoter. Primers were designed to clone S6 PDH gene promoter 5 ’end of the four deletions. And using Gateway technology to build a stable S6PDHp-GUS gene deletions and fusion expression vectors, by transformation of Arabidopsis thaliana, obtained after the T3 generation of homozygous transgenic Arabidopsis, After obtaining of T3 homozygous transgenic Arabidopsis ABA and its treatment at 4 ℃. By sampling at different time points to analyze which different pieces S6 PDHp reaction under stress circumstances of each period. The results found that the promoter sequence-836 to-236 portion has the highest low temperature response. In the ABA treatment under various transgenic plants GUS protein activity in expression patterns inconsistent performance. Promoter deletions T2396 is stress early and late in apparent response, stress response mid obvious. Promoter deletions T1396 and T836 in the early and mid-term stress response is most obvious stress late restored to its original level. In addition to the promoter deletions T236 has a response early in treatment, the rest period is not responding.3. The S6PDHp-GUS fusion expression vectors were transformed tomatoes. And GUS staining and GUS protein activity assay various tissues and organs of transgenic tomato plants were positive. It found that the promoters have different levels of activity in other tissues and organs in addition to roots. And, GUS staining and GUS activity analysis indicate that the transgenic tomato stem the strongest GUS activity. This shows that the promoter has the characteristics of tissue-specific expression. By GUS protein activity assay, indicating S6 PDHp weakest part of the active older leaves. This shows that the promoter in plant senescence process, and its expression is reduced.