橡胶树产胶生物学研究进展

天然橡胶（顺式-1,4-聚异戊二烯）是一种重要的工业原料,主要来自橡胶树。以天然橡胶的生物合成与产量形成为主要内容的产胶生物学研究为橡胶树高产遗传改良提供理论指导,近10年取得了重要进展。本文从橡胶树基因组测序、橡胶转移酶、转基因、以及转录组和蛋白质组等4个方面介绍产胶生物学研究主要进展,并讨论了相关领域研究存在的问题,对未来5~10年需重点关注的研究内容提出了建议。在介绍橡胶转移酶时,同时概述其他几种产胶植物的相关研究进展。     

Effects of phenobarbitone on serum biochemical tests in dogs

OBJECTIVES: To investigate effects of phenobarbitone on serum activities of alanine aminotransferase, alkaline phosphatase and gamma-glutamyl transferase and concentrations of bilirubin, albumin, cholesterol and total protein in dogs. ANIMALS: Ten crossbreed experimental dogs and 10 client-owned dogs of mixed breeds treated chronically with phenobarbitone to control seizures. PROCEDURES: Experimental dogs were allocated to treatment (6 mg/kg oral phenobarbitone, n = 6) and control (no treatment, n = 4) groups in which serum biochemical tests were performed at intervals during a 3-month period. Biochemical tests were performed once on the 10 epileptic dogs. RESULTS: Phenobarbitone caused increased serum alkaline phosphatase activity but did not affect gamma-glutamyl transferase activity or bilirubin, cholesterol, albumin and total protein concentrations. Phenobarbitone had minimal effect on alanine aminotransferase activity. CONCLUSIONS: Individual dogs treated with phenobarbitone may have small increases in serum alanine aminotransferase activity and variable increases in alkaline phosphatase activity but are unlikely to have alterations in gamma-glutamyl transferase activity or bilirubin, cholesterol, albumin or total protein concentrations.     

Weak activity of UDP-glucuronosyltransferase toward Bisphenol analogs in mouse perinatal development

Bisphenol A (BPA) is a widely used industrial chemical that disrupts endocrine function. BPA is an endocrine disrupting chemical (EDC) that has been demonstrated to affect reproductive organ development, brain development, metabolic disease and post-natal behavior. Accordingly, Bisphenol analogs, Bisphenol F (BPF, bis (4-hydroxyphenyl) methane) and Bisphenol AF (BPAF, 4,4-hexafluoroisopropylidene) diphenol) are used as replacements for BPA. BPA is mainly metabolized by UDP-glucuronosyltransferase (UGT), UGT2B1, but this effective metabolizing system is weak in the fetus. In the present study, we demonstrated that hepatic UGT activity toward BPAF was very weak, in comparison with BPA and BPF, in the fetus, pups and dams. Conversely, hepatic UGT activity toward BPF was very weak in the fetus and newborn pups, and was increased to the same level as BPA post-partum. In conclusion, BPAF possibly tends to accumulate in the fetus, because of weak metabolism during the perinatal period, suggesting that the metabolism of individual Bisphenol analogs requires assessment to properly gauge their risks.     

异戊烯基转移酶基因导入番茄及转基因植株再生

通过根癌农杆菌（Ａgrobacteriens）介导，利用特异启动子Ｐsag12与异戊烯基转移酶（Ｉsopenyl transferase，ipt）基因构建的嵌合基因Ｐsag12－ipt对５个番茄品种进行遗传转化，结果共获得来自３１个不同外植体的４４株转化再生植株。感染子叶的分化再生及转化植株田间生长正常。对其中部分转化再生植株进行ＰＣＲ检测证明为转基因植株，并且在田间表现出生长旺盛及抗叶片衰老的特性。     

Effects of the aglycone of ascaulitoxin on amino acid metabolism in Lemna paucicostata

Ascaulitoxin and its aglycone (2,4,7-triamino-5-hydroxyoctanoic acid, CAS 212268-55-8) are potent phytotoxins produced by Ascochyta caulina, a plant pathogen being developed for biocontrol of weeds. The mode of action of this non-protein amino acid was studied on Lemna paucicostata. Ascaulitoxin is a potent growth inhibitor, with an I₅₀ for growth of less than 1 μM, almost completely inhibiting growth at about 3 μM. Its action is slow, starting with growth inhibition, followed by darker green fronds, and then chlorosis and death. Most amino acids, including non-toxic non-protein amino acids, reversed the effect of the toxin when supplemented in the same medium. Supplemental sucrose slightly increased the activity. d-Amino acids were equally good inhibitors of ascaulitoxin activity, indicating the amino acid effects may not be due to inhibition of amino acid synthesis. Oxaloacetate, the immediate precursor of aspartate, also reversed the activity. LC-MS did not detect interaction of the compound with lysine, an amino acid that strongly reversed the effect of the phytotoxin. Metabolite profiling revealed that the toxin caused distinct changes in amino acids. Reduction in alanine, paralleled by enhanced levels of the branched chain amino acids valine, leucine and isoleucine and nearly unchanged levels of pyruvate, might indicate that the conversion of pyruvate to alanine is affected by ascaulitoxin aglycone. In addition, reduced levels of glutamate/glutamine and aspartate/asparagine might suggest that synthesis and interconversion reactions of these amino group donors are affected. However, neither alanine aminotransferase nor alanine: glyoxylate aminotransferase were inhibited by the toxin in vitro. Our observations might be explained by three hypotheses: (1) the toxin inhibits one or more aminotransferases not examined, (2) ascaulitoxin aglycone affects amino acid transporters, (3) ascaulitoxin aglycone is a protoxin that is converted in vivo to an aminotransferase inhibitor.     

Induction effects of host plants on insecticide susceptibility and detoxification enzymes of Bemisia tabaci (Hemiptera: Aleyrodidae)

BACKGROUND: The polyphagous B‐biotype Bemisia tabaci (Gennadius) has developed a high resistance to commonly used insecticides in China. To illustrate the induced changes by host plant, bioassay and biochemical research on five different host populations were investigated. RESULTS: Except for bifenthrin, all tested insecticides showed lower toxicity to the B. tabaci poinsettia population compared with other host populations. Moreover, four insecticides, the exceptions being abamectin and fipronil, showed highest toxicity towards the tomato population. The LC₅₀ values of the poinsettia population, particularly towards acetamiprid, were 14.8‐, 10.3‐ and 7.29‐fold higher than those of tomato, cucumber and cabbage respectively. The CarE activities of B. tabaci cabbage and cucumber populations were all significantly higher than those of poinsettia, cotton and tomato populations. The ratio of the cabbage population was 1.97‐, 1.79‐ and 1.30‐fold higher than that of poinsettia, cotton and tomato respectively. The frequency profiles for this activity also have obvious differences. The GST and P450 activities of the cucumber population were the lowest in the five host populations. CONCLUSION: Long‐term induction of host plants for B‐biotype B. tabaci could influence their susceptibilities to several insecticides. Rational selection and usage of insecticides for particular hosts will be helpful for resistance management and control of this species. Copyright © 2010 Society of Chemical Industry     

Cloning and characterization of two glutathione S-transferases from pyrethroid-resistant Culex pipiens

BACKGROUND: The Marin strain of Culex pipiens Say is a pyrethroid‐resistant population that was collected in Marin County, California, in 2001 and subsequently maintained in the laboratory under regular permethrin exposure. RESULTS: In this study, two cDNAs, CpGSTd1 and CpGSTd2, encoding glutathione S‐transferase (GST) were cloned from Cx. pipiens Marin. Phylogenetic analysis of the deduced amino acid sequences, CpGSTD1 and CpGSTD2, of these genes indicated that they belong to the Delta class of insect GSTs. The nucleotide and deduced amino acid sequences of CpGSTd1 and CpGSTd2 were 59 and 48% identical respectively. CpGSTD1 and CpGSTD2 were expressed in Escherichia coli and purified by affinity chromatography. The recombinant GSTs exhibited unique selectivity towards the general GST substrates 1‐chloro‐2,4‐dinitrobenzene (CDNB) and 1,2‐dichloro‐4‐nitrobenzene (DCNB), and also differed in their sensitivity to known inhibitors of GSTs. CpGSTD1 exhibited peroxidase activity with cumene hydroperoxide, while CpGSTD2 appeared to lack this activity. CpGSTD1 was able to metabolize 1,1,1‐trichloro‐2,2‐bis(4‐chlorophenyl)ethane (DDT), while DDT metabolism by CpGSTD2 was not detectable. CpGSTD1 and CpGSTD2 showed no detectable metabolism of permethrin. Gene expression of CpGSTd1 and CpGSTd2 in Marin mosquitoes was elevated about twofold in comparison with that found in a pyrethroid‐sensitive mosquito strain. CONCLUSION: The results indicate that CpGSTD1 and CpGSTD2 have unique biochemical characteristics, but they do not appear to play major roles in permethrin resistance in Marin mosquitoes. Copyright © 2012 Society of Chemical Industry     

油松谷胱苷肽转移酶Tau1单体结构稳定性研究

[目的]探究油松PtGSTU1结构与功能的关系,探讨油松PtGSTU1蛋白的单体稳定性。[方法]利用同源建模模拟PtGSTU1的三维结构,推测其N末端18位精氨酸(Arg18)和C末端103位天冬氨酸(Asp103)能够形成氢键来稳定蛋白单体结构。利用定点突变,分别将Arg18和Asp103突变为具有不同极性和构象的氨基酸残基,检测其蛋白的催化活性及结构稳定性。[结果]6个Arg18突变体均无法获得高纯度的具有正确折叠的可溶蛋白,而Asp103突变体可以表达为可溶蛋白,Asp103突变体对不同底物的催化活性和亲和力明显低于野生型,对经典底物CDNB和GSH反应的催化速率(Vmax)降低了至少9倍,对底物的催化效率(kcat/Km)也明显降低。[结论]Arg18和Asp103之间形成的氢键对稳定PtGSTU1单体结构具有重要作用,由于植物GST蛋白N端的保守性和C端结构域的多变性,Arg18的突变对结构和活性的影响大于Asp103,同时预示着C端结构域中可能存在其他氨基酸位点能够与18位精氨酸形成氢键,从而稳定蛋白单体折叠结构。