盐爪爪耐盐相关基因的cDNA-AFLP分析

以不同浓度NaCl(0、100、200、300、400和500 mmol/L)处理生长了4周的盐爪爪(Kalidium foliatum),48h后取其幼嫩叶片用于RNA提取,并进行了cDNA-AFLP表达差异分析.结果表明,用256对引物扩增出连续性差异片段155条,按照NaCl浓度升高的顺序,划分为8种差异带类型,即强→弱"、弱→强"、有→无"、无→有"、有→无→有"、无→有→无"、强→弱→强"和弱→强→弱".测序后获得176个新表达序列标签(ESTs),已将长度大于100 bp的162个ESTs登录到GenBank,登录号为HO205290~HO205450.经BLAST序列比对分析发现,盐爪爪的耐盐性可能与能量、新陈代谢、防御、转运、转录和翻译等相关蛋白有关.     

一种测定女贞子中齐墩果酸和熊果酸的新方法

为建立一种加速溶剂萃取-毛细管区带电泳测定女贞子中齐墩果酸和熊果酸的新方法.考察了萃取温度、萃取时间和萃取次数对目标物萃取效率的影响,并考察了硼砂浓度,β-环糊精浓度、pH及甲醇浓度对目标物分离的影响.结果表明:(1)萃取温度和萃取次数影响目标物的萃取效率,而萃取时间的影响很小;萃取压力影响萃取过程的重现性.(2)优化的萃取条件为:萃取压力6.9 MPa,萃取温度100℃,萃取时间5 min和萃取次数2次.(3)优化的缓冲体系为:40 mmol/L硼砂,1 mmol/Lβ-环糊精,pH 9.5及6%甲醇.(4)齐墩果酸和熊果酸分别在10~200、10~160 mg/L范围内线性良好,相关系数均大于0.996,检测限分别为3.2 mg/L和3.0 mg/L,加标回收率为93%~97%.对比了加速溶剂萃取、索氏提取以及超声提取的提取效率.(5)不同提取方法比较结果表明,加速溶剂萃取的提取效率与索氏提取法接近,但高于超声提取;加速溶剂萃取法的主要优点是消耗提取溶剂量少,提取时间短,样品用量小.     

秦岭宁陕县森林植被碳储量与碳密度特征

以秦岭南坡中段宁陕县林区2003年二类森林调查资料为基础,采用政府间气候变化委员会(IPCC)推荐使用的森林碳储量估算方法,从森林类型、林种、年龄和林分起源的角度,对该林区森林植被碳储量和碳密度进行估算。结果显示:(1)宁陕县森林植被碳储量为12.31Tg(1Tg=1×1012 g),平均碳密度为66.36Mg/hm2(1Mg=1×106 g),其各乡镇森林植被碳储量和碳密度在空间上的分布不平衡。(2)各森林类型中针叶林总碳储量为0.71Tg,平均碳密度为64.11 Mg/hm2,阔叶林总碳储量为11.61Tg,占宁陕县总碳储量的94.3%,碳密度为67.65Mg/hm2。(3)各林种中防护林碳储量最大(8.13Tg),占宁陕县总碳储量的66%,特种用途林碳密度最大(81.43Mg/hm2)。(4)不同林分起源中,天然林碳储量为12.231Tg,占宁陕县总碳储量的99.3%,人工林碳储量较小。(5)不同年龄森林中未成熟森林(包括幼龄林、中龄林和近熟林)碳储量为12.13Tg,占总碳储量的98.5%,近熟林碳密度最大(80.14Mg/hm2),幼龄林碳密度最小(39.85Mg/hm2)。研究表明,宁陕县森林具有较大的固碳能力和固碳潜力,其森林面积和蓄积是决定森林碳储量大小的重要因子,而森林碳密度的大小与森林类型、年龄组成和林分起源方式密切相关。     

不同经营措施对黄龙山辽东栎林的影响

以黄龙山林区蔡家川林场的辽东栎群落为研究对象,以未采伐为对照,采取皆伐、间伐30%、间伐15%三种经营措施,在进行自然恢复6年后,对辽东栎林建群种径级结构、群落物种多样性、土壤养分水分状况进行了对比分析,以探讨不同经营措施对黄土高原南部辽东栎(Quercus liaotungensis)林群落多样性的影响.结果表明:不同经营措施实施后建群种径级结构、群落物种多样性、土壤养分水分等指标均与未采伐林地有差异;经过间伐30%的林地,建群种径级结构、群落物种多样性、土壤养分水分等指标均有显著增加;间伐15%林地上述指标均有增加,但不显著;皆伐后的林地相对未采伐林地,建群种径级结构、群落物种多样性、土壤养分水分等指标降低.间伐与皆伐措施后比较,间伐措施的林地建群种个体生长发育良好,林下有大量幼苗,林地物种多样性丰富,土壤养分水分相对优越;皆伐措施的辽东栎建群种个体发育不良,缺乏足够数量的幼苗,林分物种多样性、土壤水分养分相对较差.未来该地区辽东栎林经营管理中,应以近自然经营措施为主,尽量减少对灌草层破坏,完善群落复层结构;间伐强度拟定为30%,尽量间伐劣质木、病虫木,促进群落向异龄方向发展.     

波形蛋白介导猪繁殖与呼吸综合征病毒感染Marc-145细胞作用的初步研究

为了研究波形蛋白在介导PRRSV感染过程中的作用,根据GenBank中已发表的波形蛋白序列,设计特异性引物,利用RT-PCR方法从Marc-145细胞中扩增目的基因,克隆入pET-28a,转化表达菌BL21(DE3)中进行诱导表达,用SDS-PAGE和Western blot鉴定表达产物,将表达产物纯化后免疫BALB/c小鼠制备血清,用病毒阻断实验验证PRRSV与波形蛋白及其抗体的关系。用ELISA方法确定了重组波形蛋白与PRRSV结构蛋白N蛋白和囊膜蛋白GP5蛋白之间的关系。结果表明,成功的从Marc-145细胞中扩增出全长的波形蛋白基因,将其克隆入pET-28a,诱导后得到高效表达,表达蛋白纯化后免疫小鼠抗体效价达到105,病毒阻断实验表明波形蛋白能部分阻断PRRSV感染,其抗血清能完全阻断PRRSV感染。ELISA结果表明波形蛋白能与N蛋白结合而不与GP5蛋白结合。此结果为PRRSV感染的细胞的受体机制增添了新的内容,为PRRSV感染过程中受体间的相互作用关系研究奠定基础。     

沙地生境和平茬年限对沙柳叶功能特征的影响

研究了毛乌素沙地南缘水滨边丘间地和干旱丘顶两种生境下沙柳平茬后不同年限(1、2、3～4和5～6年)对沙柳叶片光合气体交换、水分利用效率、结构特性及氮、磷养分含量等叶功能特征的影响.结果表明:水滨边丘间地沙柳叶片的净光合速率、气孔导度、瞬时和长期水分利用效率均高于丘顶,但N、P养分含量低于丘顶；两种生境下沙柳的叶结构性状无显著差异.干旱生境下沙柳主要通过增加养分含量、降低光合和水分利用来生存.随平茬年限增加,叶片净光合速率和气孔导度显著下降,氮含量和瞬时水分利用效率亦呈下降趋势,且二者之间呈显著正相关；平茬后1年的叶面积最高,比叶质量和叶干物质含量最低,此后比叶质量增加,叶面积和干物质含量不变；叶结构性状与光合及养分特性之间无显著相关性.叶光合活性和氮含量的下降是沙柳随平茬年限增加而衰败的重要原因.     

<Emphasis Type="Italic">Rhizobium sphaerophysae</Emphasis> sp. nov., a novel species isolated from root nodules of <Emphasis Type="Italic">Sphaerophysa salsula</Emphasis> in China

Four gram-negative, aerobic, motile, non-spore, forming rods with a wide pH and temperature range for growth (pH 7.0–11.0, optimum pH 8.0; 20–45°C, optimum 28°C) strains were isolated from root nodules of Sphaerophysa salsula and characterized by means of a polyphasic approach. Phylogenetic analysis based on 16S rRNA gene sequences revealed that the four strains formed a new lineage related to the genus Rhizobium and the sequence similarities between the isolate and the most related type strain Rhizobium giardinii was 96.5%. These strains also formed a distinctive group from the reference strains for defined Rhizobium species based on housekeeping gene sequences (atpD and recA), BOX-PCR fingerprinting, phenotypic features and symbiotic properties. The representative strain CCNWGS0238^T has DNA-DNA relatedness of less than 33.4% with the most closely related species R. giardinii. It is therefore proposed as a new species, Rhizobium sphaerophysae sp. nov., with isolate CCNWGS0238^T (=ACCC17498^T = HAMBI3074^T) as the type strain.     

Methylobacterium soli sp. nov. a methanol-utilizing bacterium isolated from the forest soil

A Gram-negative, pink-pigmented, non-spore-forming rod shaped, methanol-utilizing bacterium, strain YIM 48816(T), was isolated from forest soil collected from Sichuan province, China. Strain YIM 48816(T) can grow at 4-37 °C, pH 5.0-7.0 and 0% NaCl (w/v). Based on 16S rRNA gene sequence similarity studies, it belonged to the genus Methylobacterium, and formed a phyletic line. The 16S rRNA gene sequence similarities were 96.2% to Methylobacterium mesophilicum DSM 1708(T) and 96.0% to Methylobacterium brachiatum DSM 19569(T), and the phylogenetic similarities to all other Methylobacterium species with validly published names were less than 96.0%. The major menaquinones detected were Q-10 (97.14%) and Q-9 (2.86%). The major fatty acids were C18:1 ω7c (80.84%). The DNA G + C content was 66.2 mol%. It is apparent from the genotypic and phenotypic data that strain YIM 48816(T) belongs to a novel species of the genus Methylobacterium, for which the name Methylobacterium soli sp. nov. is proposed. The type strain is YIM 48816(T) (CCTCC AA 208027(T) = KCTC 22810(T)).     

Petunia <Emphasis Type="Italic">AGAMOUS</Emphasis> Enhancer-Derived Chimeric Promoters Specify a Carpel-, Stamen-, and Petal-Specific Expression Pattern Sufficient for Engineering Male and Female Sterility in Tobacco

Previous studies have shown that the AtAGIP promoter derived from the Arabidopsis AGAMOUS (AG) second intron/enhancer specifies a carpel- and stamen-specific expression in its native host species, but not in heterologous species such as tobacco, which restricts its application in the engineering of male and female sterility. These findings also imply that the AG regulatory mechanism that has evolved in Arabidopsis may, to some extent, have diverged from that of tobacco. To test whether a similar chimeric promoter created using the AG second intron/enhancer can overcome this barrier of evolutionary divergence in closely related species, we generated forward- and reverse-oriented chimeric promoters, fPtAGIP and rPtAGIP, from the petunia AG second intron/enhancer (PtAGI) fragment and tested them in tobacco, which, like petunia, belongs to the Solanaceae family. Our results demonstrate that both fPtAGIP and rPtAGIP confer similar carpel- and stamen-specific expression without any leaky activity in vegetative tissues in tobacco as revealed by tissue-specific gene expression and tissue ablation. This pattern resembles that driven by the AtAGIP in Arabidopsis and indicates that the AG regulatory mechanism is more conserved between tobacco and petunia than between tobacco and Arabidopsis. The petunia-derived promoters also exhibited petal-specific activity, and their activities in floral organs were substantially influenced by the orientation of the PtAGI enhancer, with reverse-oriented enhancers displaying approximately double the effectiveness of forward-oriented enhancers. These two properties are novel and have not been observed previously with AtAGIP promoters. Furthermore, we found that PtAGIP promoter-driven tissue ablation is effective for engineering complete sterility in plants, and the resulting sterile trait is stable for at least three mitotic generations at various temperature regimes, which is important for the complete containment of seed-, pollen-, and fruit-mediated gene flow in field conditions.