小麦钙调素新亚型TaCaM5的克隆及表达分析

利用RT-PCR技术,从条锈菌诱导的小麦叶片中分离出一个编码CaM基因的cDNA序列, 经氨基酸序列分析确定其为一个新的小麦CaM亚型,暂被命名为TaCaM5。TaCaM5包含一个完整450 bp的开放阅读框,编码149个氨基酸;编码的蛋白不含跨膜区、无信号肽、定位在胞内,具有4个EF-hand保守结构域。在目前已知的CaM基因中,TaCaM5与玉米CaM基因的亲缘关系最近,相似性高达97%。该基因在根、茎、叶等组织中均有不同程度的表达;并且受条锈菌诱导表达,在非亲和组合与亲和组合中,分别在接种后6 h和24 h表达量最高。外源植物激素脱落酸、茉莉酸甲酯和乙烯诱导TaCaM5上调表达,水杨酸诱导其下调表达。TaCaM5在机械伤害、干旱和低温条件下表达量上升,在高盐环境下表达量降低。表明TaCaM5可能通过茉莉酸和乙烯等信号途径参与小麦对条锈菌的防御反应,同时参与机械伤害、低温和干旱环境下的Ca²⁺-CaM信号转导途径。     

Non-destructive Limits to Seed Growth and Leaf Protease Activities in Nodulating and Non-nodulating Soybean Isolines

Leaf senescence leads to a progressive decline in the photosynthetic competence of the leaf. This paper describes some effects of source:sink imbalance on leaf protein catabolism and senescence in soybean. We manipulated pod growth by restricting 100 or 50 % (PR-100 or PR-50, respectively) of young pods at the R4 stage in plastic drinking straws. This effectively reduces final seed mass without interrupting the vascular connections of pods. Nodulating (NOD+) and non-nodulating (NOD−) isolines of the 'Clay' soybean were grown in drainage lysimeters and three pod-restriction (PR) treatments were compared. Pod restriction decreased seed biomass per plant as a result of lower individual seed mass, which was only partially balanced by the increase in seed number. The nitrogen concentration in seeds remained unchanged in NOD+ plants, while it increased with the degree of sink restriction in seeds of NOD− plants. Leaf soluble protein, CO₂ exchange rate and seed nitrogen content were consistently lower in NOD− plants; the leaf protein level remained stable with time in PR-100 plants, decreased for PR-50 and dropped for controls. Endoprotease (HBase) and carboxypeptidase (CPase) activities were significantly lower in leaves from PR-100 plants, while aminopeptidase activity was enhanced, indicating a de novo synthesis of leaf protein. This is consistent with the reported accumulation of vegetative storage proteins (VSPs) in soybean and other legumes after moderate or severe sink reduction. Thus, small modifications of the source:sink ratio such as those obtained by the non-destructive PR technique have an impact on leaf protein catabolism. Nodulating and non-nodulating soybean isolines showed similar responses to PR in terms of leaf senescence initiation and progression, but the rate of the processes appear to be largely influenced by plant N status.     

Low glyphosate rates do not affect Citrus limonia (L.) Osbeck seedlings

BACKGROUND: Glyphosate is used to control weeds in citrus orchards, and accidental spraying or wind drift onto the seedlings may cause growth arrest owing to metabolism disturbance. Two experiments were carried out to investigate the effect of non‐lethal rates (0, 180, 360 and 720 g AI ha^?1) of glyphosate on four‐month‐old ‘Cravo’ lime, Citrus limonia (L.) Osbeck, seedlings. Photosynthesis and the concentrations of shikimic acid, total free amino acids and phenolic acids were evaluated. RESULTS: Only transitory effects were observed in the contents of shikimate and total free amino acids. No visual effects were observed. CONCLUSION: The present study showed that glyphosate at non‐lethal rates, which is very usual when accidental spraying or wind drift occurs in citrus orchard, did not cause severe metabolic damage in ‘Cravo’ lime seedlings. Copyright © 2009 Society of Chemical Industry     

Development and trial of a bovine herpesvirus 1 - thymidine kinase deletion virus as a vaccine

SUMMARY An Australian bovine herpesvirus 1 (BHV1) isolate with a defined (427 base pair) deletion in the protein coding region of the thymidine kinase gene was obtained by standard marker rescue procedures. After selection in the presence of the nucleotide analogue 5iodo-deoxy-uridine the virus was analysed by hybridisation with three differential oligonucleotide probes, restriction endonuclease profile studies and DNA sequence analysis. The virus elicited an immune response in recipient animals after either intramuscular or intravenous administration and produced no significant deleterious side-effects when administered at a dose sufficient to stimulate the host immune response. The safety and immunogenicity of the recombinant BHV1 virus 39B1 were similar to those reported for other registered BHV1 vaccines and the virus would appear to be suitable for the production of a vaccine seed lot and more exhaustive field trials as a prelude to commercial vaccine production and registration.     

Ovum Pick-up in Sheep: a Comparison between Different Aspiration Devices for Optimal Oocyte Retrieval

In vivo ovum pick-up (OPU) in sheep may be improved with a proper choice of aspiration elements (needle and tubing) and aspiration vacuum pressure. In the present study, two experiments were carried out. In Expt 1, visible follicles in ovaries of slaughtered ewes (treated separately according to their diameters: small<3 mm, medium 3-5 mm and large>5 mm) were aspirated using different combinations of the three studied factors such as aspiration flow rate (10, 20, 30, 40 and 50 ml water/min), needle gauge (18 and 20 G) and tubing inner diameter (1, 2 or 3 mm internal diameter). In Expt 2, a study with two 18 G needles of different lengths (18 G: 82 mm; 18 GL: 600 mm) was carried out, using ovaries obtained post-mortem, and performing in vivo laparoscopic follicular aspiration on ewes. We considered good quality oocytes as those with both complete compact cumulus and a homogeneous cytoplasm. Recovery rate, proportion of good quality oocytes (good quality oocytes/100 oocytes recovered) and overall efficiency (good quality oocytes/100 follicles aspirated) were noted. In Expt 1, aspiration flow rate affect remarkable proportion of good quality oocytes (69.5%, 50.5%, 44.8%, 36.5% and 28.3% for flows from 10 to 50 ml/min respectively, p<0.05). Needle gauge did not affect aspiration device efficiency. Thin and intermediate tubings were more effective (overall efficiency rates: 34.9%, 32.3% and 28.1% for 1, 2 and 3 mm respectively, p<0.05). Follicle size did not affect recovery rate, but proportion of good quality oocytes was higher for large (77.9%) and medium (64.4%) follicles (p<0.05). Finally, some combinations of the aspiration device showed greater effectiveness. In Expt 2, needle length did not influence recovery rate, but good quality oocytes rate was significantly modified both post-mortem and in vivo (good quality rate for 18 G vs 18 GL needles: 69.5% vs 47.7% and 58.1% vs 25.4%, post-mortem and in vivo respectively, p<0.05). We conclude that low-aspiration flow rates (10 and 20 ml/min) with thin or intermediate tubings (1 and 2 mm), and any short needle (18 G or 20 G) are the most adequate aspiration factors for OPU in sheep.     

Involvement of glucagon-like peptide 1 in the glucose homeostasis regulation in obese and pituitary-dependent hyperadrenocorticism affected dogs

The incretin glucagon-like peptide 1 (GLP-1) enhances insulin secretion. The aim of this study was to assess GLP-1, glucose and insulin concentrations, Homeostatic Model Assessment (HOMA_{insulin sensitivity} and HOMA_{β-cell function}) in dogs with pituitary-dependent hyperadrenocorticism (PDH), and compare these values with those in normal and obese dogs. The Oral Glucose Tolerance Test was performed and the glucose, GLP-1 and insulin concentrations were evaluated at baseline, and after 15, 30, 60 and 120 minutes. Both basal concentration and those corresponding to the subsequent times, for glucose, GLP-1 and insulin, were statistically elevated in PDH dogs compared to the other groups. Insulin followed a similar behaviour together with variations of GLP-1. HOMA_{insulin sensitivity} was statistically decreased and HOMA_{β-cell function} increased in dogs with PDH. The higher concentrations of GLP-1 in PDH could play an important role in the impairment of pancreatic β-cells thus predisposing to diabetes mellitus.