A target-site mutation is present in a glyphosate-resistant Lolium rigidum population

Annual ryegrass (Lolium rigidum) is the only weed species to have evolved resistance to the broad‐spectrum herbicide glyphosate in Australia. A population that had failed to be controlled by glyphosate was collected from a vineyard in the Adelaide Hills region of South Australia. Dose–response experiments on this population (SLR 77) showed that it was glyphosate resistant, with an LD₅₀ that was 1.9–3.4 times higher than that of a susceptible population (VLR 1). The movement of radiolabelled glyphosate within SLR 77 plants showed that this population did not have the differential glyphosate translocation mechanism of resistance common to several other Australian glyphosate‐resistant populations. Subsequent analysis of shikimic acid accumulation within the plant after glyphosate treatment showed that this population accumulated significantly less shikimic acid than a susceptible population, but more than a glyphosate‐resistant population with the translocation mechanism, indicating the possible involvement of another mechanism of resistance. Sequencing of a portion of the SLR 77 5‐enolpyruvylshikimate‐3‐phosphate synthase gene was carried out and a mutation causing an amino acid change at position 106 from proline to threonine was identified. This mutation is likely to be responsible for glyphosate resistance in this population, as mutations in this position have been found to be responsible for glyphosate resistance in goosegrass (Eleusine indica) from Malaysia. This paper represents the first report of target‐site glyphosate resistance in L. rigidum and provides evidence that this species has at least two mechanisms of glyphosate resistance present in Australia.     

Influence of transection of the cervical sympathetic trunk on content of NO and expression of NOS in placenta of rats with pregnancy-induced hypertension syndrome

AIM:To observe the influence of transection of the cervical sympathetic track (TCST) on the content of NO and the expression of eNOS mRNA and iNOS mRNA in placenta of the rats with pregnancy-induced hypertension syndrome (PIH).METHODS: Pregnant Wistar rats were randomly divided into 5 groups: control group (C): saline was injected subcutaneously from 14th day to 20th day of gestation;PIH group 1 (H1) and group 2 (H2): L-NAME was respectively injected with 125 mg/kg and 62.5 mg/kg,respectively,then the other procedures were the same as group C;Operation group (O): TCST was operated on 14th day of the gestation,then the other procedures were the same as group H1;sham operation group (S): the cervical sympathetic trunk was only separated and exposed on 14th day of the gestation,then the other procedures were the same as group H1.RESULTS: (1) Except the base value of the BP and protein in urine of the pregnant rats,all the parameters observed in group H1 and H2 were higher than those in group C significantly (P<0.01),and in group H2 were lower than those in group B1 markedly (P<0.01).(2) In comparison with those in group C,the size and body weigh of fetus in group H1,H2 decreased markedly (P<0.01).The above indexes in group H1 were lower than those in group H2 markedly (P<0.01,P<0.05).The changes of the rate of embryo absorption and fetal death,and deformity rate of the fetal rats were contrary to the above indexes.(3) The content of NO and the expression of eNOS mRNA and iNOS mRNA in placenta in group H1 and H2 were lower than those in group C markedly (P<0.01).Those in group H1 were lower than those in group H2 obviously (P<0.01,P<0.05).Those in group O were higher than those in group H1 markedly (P<0.01).CONCLUSION: TCST protects pregnant rats against PIH,and it was related to the mRNA expression of eNOS and iNOS and the content of NO in placenta tissue.     

果实特异启动子调控薄皮甜瓜ACC合成酶cDNA反义表达载体的构建

应用反义RNA技术抑制甜瓜成熟过程中内源乙烯的合成,从而培育耐贮运品种是解决甜瓜延熟保鲜难题的可行新方法。根据GenBank中甜瓜、黄瓜ACC合成酶基因氨基酸保守序列设计引物,从成熟的薄皮甜瓜(齐甜1号)果肉组织中提取总RNA,经RT-PCR扩增得到约0.7kb的ACC合成酶cDNA片段,将其克隆到质粒载体pGEM-TEasy中,测序表明,该基因为777bp,编码258个氨基酸;从番茄(东农706)叶片组织中提取总DNA,经PCR扩增得到约2.2kb的E8基因片段,将其克隆到质粒载体pGEM-TEasy中,测序表明,该基因为2192bp;以pCAM2301为起始植物表达载体,pCAM-GT为中间载体,成功构建了果实特异启动子(E8)调控薄皮甜瓜ACC合成酶cDNA反义表达载体,采用冻融法将其转入根癌农杆菌LBA4404,得到了完整的Ti质粒表达载体系统。     

Pineapple organic acid metabolism and accumulation during fruit development

Developmental changes in pineapple (Ananas Comosus (L.) Merrill) fruit acidity was determined for a ‘Smooth Cayenne’ high acid clone PRI#36-21 and a low acid clone PRI#63-555. The high acid clone gradually increased in fruit acidity from 1.4 meq/100 ml 6 weeks from flowering, and peaked a week before harvest at ca 10 meq/100 ml. In contrast, the low acid clone increased in acidity 6 to 8 weeks after flowering, peaked 15 weeks after flowering at ca. 9 meq per/100 ml and then sharply declined in 2 weeks to 6 meq/100 ml. The increased in total soluble solids (TSS) of the low acid clone began 6 weeks after flowering and for the high acid clone at 12 weeks after flowering. The increase in titratable fruit acidity (TA) paralleled the changes in the citric acid content of both clones. Citric acid content increased from less than 1 mg/g at 6 weeks after flowering to 6 to 7 mg/g, 9 weeks later. The malic acid concentration in both clones varied between 3 and 5 mg/g and showed no marked changes just before harvest. The developmental changes in fruit potassium were significantly correlated with fruit acidity and fruit total soluble solids in both the high and low acid clones. Developmental changes in acid-related enzymatic activities showed an increase in citrate synthase (EC 4.1.3.7) activity that occurred a week before harvest, coincided with the peak in citric acid in the high acid clone. An increase in aconitase (ACO, EC 4.2.1.3) activity was observed just before harvest as the decline in acidity occurred in the low acid clone. The activities of phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31), malate dehydrogenase (MDH, EC 1.1.1.37) and malic enzyme (ME, EC 1.1.1.40) did not parallel any changes in fruit acidity. The results indicated that the change in pineapple fruit acidity during development was due to changes in citric acid content. The major difference in acid accumulation occurred in the low acid clone just before harvest when acidity declined by one-third. The activities of citrate synthase and aconitase possibly played a major role in pineapple fruit acidity changes.     

Renoprotective effects of sesamin on renal hypertensive and hyperlipidemic rats

AIM: To investigate the effects of sesamin on progression of renal injury in renal hypertensive and hyperlipidemic rats (RHHR). METHODS: RHHR was induced by 2K1C and high lipid baitvessel. After 7 weeks of intragastric administration with sesamin, the contents of serum creatinine (Scr), blood urea nitrogen (BUN), 24 h urinary protein excretion (UPE) were measured. In addition, the activity of total antioxidative capacity (T-AOC), superoxide dismutase (SOD), the concentrations of malondialdehyde (MDA), hydrogen peroxide (H2O2), and the nitric oxide synthase (NOS) and nitric oxide (NO) levels in renal homogenate were measured. RESULTS: Compared with the model group, seasamin (in 100 mg·kg-1 and 33 mg·kg-1 groups) evidently decreased the contents of Scr, BUN, UP and the concentration of MDA, iNOS, H2O2 in renal tissure. It also improved the levels of NO, cNOS and activity of SOD, T-AOC in renal tissure. CONCLUSION: Sesamin ameliorates hypertensive and hyperlipidemic-induced renal injury, probably by enhancing antioxidative activity, scavenging hydroxyl radical and restraining iNOS level.     

香蕉柠檬酸合酶基因MaGCS的克隆及生物信息学分析

采用RACE-PCR的方法首次在香蕉中获得一个柠檬酸合酶基因的cDNA序列。经序列测定和Blastx比对分析表明，该cDNA全长1 885bp，编码513个氨基酸残基，具有植物柠檬酸合酶基因的特征结构域，并与其他植物来源的乙醛酸循环体的柠檬酸合酶具有很高的序列相似性，将其命名为MaGCS(Musa glyoxysomal citrate synthase)，并对其进行生物信息学分析，初步预测其理化性质及功能等。     

小麦淀粉合酶基因I的克隆及反义和RNAi载体的构建

采用RT-PCR方法从小麦品种豫教2号发育的籽粒中克隆出淀粉合酶I基因(starch synthase I,SSI)部分cDNA片段(795 bp)(GenBank No.EF221762),同源性比较结果显示,它与GenBank上已报道的SSI基因有高度同源性。以p WM101质粒为基础,构建了由35S启动子调控的SS I基因的反义表达载体p WM101SSI;另外,还以pFGC5941质粒为基础,构建了SSI基因的RNAi载体pFGC5941SSIsa,这些载体的构建为研究此基因的功能奠定了基础。     

日本血吸虫GSK3蛋白编码cDNA的克隆、表达及其重组蛋白的免疫保护研究

本研究利用生物信息学分析了日本血吸虫(Schistosoma japonicum,sj)Glycogen synthase kinase 3(GSK3)蛋白,并对其中一个GSK3蛋白的编码cDNA进行了克隆和原核表达,制备了特异性的多克隆抗体.同时,还初步评估了重组蛋白的免疫保护效果.生物信息学分析表明在日本血吸虫数据库存在两种GSK3蛋白,且其中一个SjGSK3在日本血吸虫不同发育时期均有转录.Western blot结果表明本研究制备的抗体能特异性识别日本血吸虫SjGSK3重组蛋白,表明该重组蛋白具有良好的免疫原性.动物实验表明免疫SjGSK3重组蛋白的动物与佐剂对照组比较分别获得了平均10.6％减虫率和40.5％肝脏减卵率.