不同碳源氮源对腐霉菌株生长的影响

在喀麦隆芋芳根涡病病原研究中，测定了不同碳源、氮源对分离自喀麦隆芋艿田间及来自中国的腐霉菌株生长习性的影响。实验结果表明，除了钟腐霉（Ｐｙｔｈｉｕｍｖｅｘａｎｓ）菌株只能吸收葡萄糖、果糠而不能吸收蔗糖外，其余腐霉菌株均能利葡萄糖、果糖、蔗糖作为碳源。所有试验菌株均不能利用半乳糖、麦芽糖及可溶性淀粉。钟腐霉菌株之形态特征与特殊碳源营养的相关性显示了用碳源作为菌株鉴定辅助标准的可行性，氮源营养试验结果表明，所有腐霉菌株均能利用四种氮源，根据不同氮源对菌丝生长的影响，依次为Ｌ－天门冬氨酸，Ｌ－精氨酸，硝酸铵，硝酸钾。     

Storage of Non-structural Carbohydrates in Sweet Sorghum [Sorghum bicolor (L.) Moench.]: Comparison of Sterile and Fertile Lines

The accumulation of soluble carbohydrates in vegetative parts of sweet sorghum ( Sorghum bicolor (L.) Moench.) represents an intermediate reserve pool for grain filling as well as an irreversible storage up to maturity. In order to study the effect of Sterility on soluble sugar accumulation in vegetative parts, two cytoplasmic male sterile (CMS) lines of sweet sorghum were compared with their corresponding fertile mamtainers for non-structural carbohydrate concentrations in shoots. Plants were harvested at four different physiological stages. The lines showed significant differences in the yield of non-structural carbohydrates. At maturity, the yields of soluble carbohydrates from vegetative parts of the two sterile lines were 93 % and 43 % higher than the yield of the two corresponding fertile lines. At that stage, sucrose represented about 80 % of total soluble carbohydrates and the higher yield of soluble carbohydrates of sterile lines was mainly due to the higher yield of sucrose. This indicates that the yield of extractable carbohydrates, especially sucrose, in vegetative parts of sweet sorghum could be considerably improved by using sterile lines which are utilized in hybrid production.     

鹿流行性出血病毒分子生物学研究进展

鹿流行性出血病毒是一种在全世界野生及驯养的有蹄动物中广泛存在的重要病原体.该病毒属呼肠病毒科环状病毒属,有12个血清型,是一种有10个节段(L1-L3、M4-M6、S7-S10)的双链RNA病毒,编码10个蛋白(VP1-7和NS1、NS2、NS3/NS3A).VP7蛋白具有很强的抗原性且保守性最高.VP2与病毒型特异性有关,可诱导产生中和抗体.VP3为群特异性抗原,高度保守,具有亲水性保守区域.非结构蛋白NS1、NS2和NS3/NS3A及其编码序列均相当保守.该病毒的分子生物学诊断技术主要有PCR和核酸探针杂交技术.     

The effect of PHA-lectins and diet composition on performance and intestinal development in weaning piglets

Anti-nutritional factors such as PHA-lectins (phytohaemagglutinin) in piglet diets can compromise piglet's performance and health by gut damage, which is especially important at weaning. Two trials were conducted to investigate the effect of PHA and fermentable carbohydrates on performance, gut morphology, physiology and microbiology in piglets weaned at 26 days.

We studied dietary PHA as a model to standardize gut damage and its effect on intestinal morphology and microbiology in piglets until 2 weeks post weaning. In general, significant lower values were observed on day 7 compared to day 14 post weaning (d7 vs. d14: 358 vs. 442 μm villus height; 42.1 vs. 108.7 U/g protein sucrase-isomaltase), indicating gut maturation. We observed very few changes between day of weaning to day 7 post weaning, lactobacilli counts being most affected (8.9, 6.9, 7.4 cfu/g for d0, d7, d14). Apart from these marked time effects the effect of PHA was negligible and thus a less suitable model to standardize gut damage in this setup.

Furthermore we studied the effect of PHA in combination with rapid or slow fermentable carbohydrates (CHO) on performance and physiological parameters. We observed a dramatic effect on performance (ADG g/d: 245 no PHA + low CHO, 111 PHA + low CHO, 132 PHA + rapid CHO, 105 PHA + slow CHO), which was less clearly reflected in the physiological parameters.     

猪繁殖与呼吸综合征病毒非结构蛋白Nsp9基因的克隆与原核表达

通过PCR方法从重组质粒pSK-B2扩增得到非结构蛋白Nsp9的基因片段。将基因克隆至原核表达载体pET-28a( ),得到重组表达载体pET-28-Nsp9,转化Escherichia coliBL21(DE3)细胞。经IPTG诱导,Nsp9蛋白以包涵体形式表达,SDS-PAGE分析表明重组蛋白的分子量约为27.3 kD,表达量占菌体蛋白的43.2%。Western-Blotting结果表明重组蛋白可被PRRSV阳性血清所识别。表达的重组蛋白为进一步研究PRRSV的免疫特性和分子生物学功能奠定了基础。     

生物炭对干旱胁迫下咖啡苗生长及非结构性碳水化合物的影响

为明确在干旱胁迫下施用生物炭对咖啡苗生长及非结构性碳水化合物（NSC）含量的影响，以正常水分处理（W1：土壤水分为最大持水量的65%~70%，不施用生物炭；W2：土壤水分为最大持水量的65%~70%，施用相当于烘干土质量5%的生物炭）为对照，设置不同生物炭处理（D1：不施生物炭；D2：施用量为干土质量的5%）下进行持续干旱与复水的盆栽试验，干旱与复水共设4个处理，分别为持续干旱9 d（DL）、持续干旱13 d（DS）、持续干旱9 d+复水后3 d（DL+R）、持续干旱13 d+复水后3 d（DS+R），分析各器官（根、茎、叶）干物质量、NSC组分含量及其质量的变化。结果表明：与正常水分处理（W1）相比，持续干旱9 d时（DL），D1处理咖啡苗叶的可溶性糖含量、淀粉含量和NSC比率分别下降22.5%、21.1%和21.1%，根的可溶性糖含量和NSC比率分别显著提高8.7%和62.8%，茎的可溶性糖含量和NSC比率分别显著提高22.0%和28.2%。持续干旱13 d时（DS），D1处理较W1处理的根、茎、叶干质量和总干物质量分别下降30.6%、22.2%、34.8%和30.8%，叶的NSC含量和NSC比率分别下降23.7%和16.4%，根的NSC含量和NSC比率分别增加33.8%和57.9%；生物炭处理（D2）较不施生物炭的处理（D1）总干质量增加16.7%，根的NSC含量和茎的NSC比率分别下降18.0%和24.1%，叶的NSC含量和NSC比率分别增加22.8%和15.0%。持续干旱9 d复水后3 d时（DL+R），生物炭处理（D2）对各器官NSC含量恢复作用显著，且与正常水分处理（W1）间无显著差异；干旱13 d复水后3 d时（DS+R），不施生物炭处理（D1）的咖啡苗生长未恢复，而生物炭处理（D2）对咖啡苗恢复作用显著，D2较D1处理的总干质量增加20.3%，叶的NSC含量增加22.7%，根和茎的NSC含量分别下降11.8%和15.3%。可见，施用生物炭是增强咖啡抗旱性和减缓咖啡NSC组分剧烈变化的有效途径。     

牦牛源BVDV非结构蛋白NS5A的原核表达及抗原表位分析

【目的】利用原核表达系统体外表达牛病毒性腹泻病毒(Bovine viral diarrhea virus,BVDV)NS5A基因,获得非结构蛋白NS5A,对其进行核苷酸、氨基酸序列分析,以解析BVDV非结构蛋白NS5A的功能。【方法】参考BVDV-1型毒株V006的NS5A基因序列(GenBank登录号:KX170647)设计并合成1对特异性引物,以分离到的牦牛BVDV GSTZ毒株cDNA为模板,PCR扩增NS5A基因片段,并克隆至表达载体pET-28a (+)中,构建重组原核表达载体pET28a-NS5A。经酶切初步鉴定及测序鉴定正确后,转化大肠杆菌BL21(DE3)感受态细胞,然后利用IPTG诱导表达。经10% SDS-PAGE电泳及Western blotting分析鉴定重组蛋白的表达,并根据NS5A基因的序列构建遗传发育进化树,利用DNAStar软件预测NS5A蛋白的亲水性、表面可塑性和抗原性等特性,并结合二级结构的预测对NS5A蛋白的B细胞抗原表位进行预测。【结果】PCR扩增NS5A目的基因片段为1 488 bp,双酶切和测序鉴定结果证明,重组质粒pET28a-NS5A构建成功。经10% SDS-PAGE电泳及Western blotting鉴定重组蛋白,表达出了大小为55 ku的目的蛋白,大小与预期结果相符。通过对不同BVDV毒株NS5A基因序列构建遗传发育进化树,显示GSTZ毒株NS5A在遗传进化特征上属于BVDV-1型。NS5A蛋白的亲水性主要位于12—21、32—69、75—113、120—135、143—147、152—163、165—180、215—230、265—274、296—340、348—378、389—447、455—463、469—495位氨基酸处,表面可塑性主要位于14—18、37—42、76—81、86—109、154—160、169—178、218—228、297—309、348—358、365—373、414—442、430—437、454—460位氨基酸处,柔性区域较多,主要位于14—21、37—43、67—82、86—93、97—110、152—158、169—179、218—231、240—255、296—310、313—328、344—359、364—373、413—422和472—483位氨基酸处。NS5A蛋白的B细胞抗原表位主要位于15—18、76—81、154—158、169—178、218—228、297—309、348—358、365—373和414—422位氨基酸处。【结论】成功表达并鉴定了牦牛源BVDV的非结构蛋白NS5A,系统发育进化树表明BVDV GSTZ株基因型属于BVDV-1型,NS5A蛋白具有良好的抗原性,为深入解析BVDV非结构蛋白NS5A的自身结构功能、免疫学特性以及进一步研究非结构蛋白对病毒复制的影响提供参考。     

Effect of water stress conditioning on morphology,physiology and field performance of Pinus halepensis Mill. seedlings

Five-month-old Pinus halepensis Mill. seedlings were subjected to 4 irrigation treatments for 8 weeks. After the treatments, morphological and physiologicalattributes assessed included height, diameter, dry weight, water relations parameters, and determination of N, P, K, soluble carbohydrates, and starch concentrations. The remaining seedlings were field planted. Survival and height growth were recorded forfour growing seasons after planting. The mostsignificant effect of irrigation was onmorphology. Increased irrigation lead toseedlings with significantly higher height,root collar diameter and shoot and rootbiomass. Starch and soluble carbohydrateconcentrations were also affected byirrigation. However, parameters derived frompressure-volume curves were not significantlydifferent among treatments, nor were N, P, andK concentrations. There were no differencesamong treatments for survival in the field,which was highly related to summer rainfall. Incomparison, absolute and relative heightgrowth showed some significant but minordifferences among treatments.     

Biomass dynamics of Erythrina lanceolata as influenced by shoot-pruning intensity in Costa Rica

Pruning of agroforestry trees, while reducing shade of the crops, usually reduces both biomass production and nitrogen fixation. Short pruning cycles are often not sustainable on the long run, because tree production declines over subsequent pruning cycles. We compared biomass and labile carbohydrate dynamics of Erythrina lanceolata Standley (Papilionaceae) shade trees under total and partial pruning regimes in a vanilla (Vanilla planifolia L.) plantation in South-western Costa Rica. The highest biomass production was measured in the unpruned control, followed by trees with 50% of the leaf pruned every three months, while total pruning every six months resulted in the lowest biomass pruduction. In the more productive treatments, a higher proportion of the production was in branches. Because, the N content of woody branches was high, they were important for nitrogen cycling. In the partial pruning treatment more nitrogen was returned to the soil from litter and woody branches than from pruned leaf. Sugar concentrations were not different between treatments and the dynamics of non-structural carbohydrates (sugar and starch) seems to depend more on plant phenology than pruning treatment. However, the starch concentrations in the total pruning were lower than in the other treatments.This revised version was published online in November 2005 with corrections to the Cover Date.     

不同种源北美香脂杨苗木碳水化合物积累差异分析

对9个不同种源的北美香脂杨在加拿大温哥华扦插繁殖的苗木枝条进行分析,分另4测定带芽枝条可溶性糖、淀粉、糖组分的含量,扦插后萌发的叶干质量和根干质量。结果表明：不同种源枝条的可溶性糖和淀粉含量差异极显著;扦插生根和叶的干质量与淀粉含量间呈现正相关关系,与各地的地理位置（纬度、经度、海拔）无显著相关性;不同种源枝条内淀粉含量与其扦插生根特性呈正相关关系。