Fine mapping a QTL qCTB7 for cold tolerance at the booting stage on rice chromosome 7 using a near-isogenic line

Low temperature at the booting stage is a serious abiotic stress in rice, and cold tolerance is a complex trait controlled by many quantitative trait loci (QTL). A QTL for cold tolerance at the booting stage in cold-tolerant near-isogenic rice line ZL1929-4 was analyzed. A total of 647 simple sequence repeat (SSR) markers distributed across 12 chromosomes were used to survey for polymorphisms between ZL1929-4 and the cold-sensitive japonica cultivar Towada, and nine were polymorphic. Single marker analysis revealed that markers on chromosome 7 were associated with cold tolerance. By interval mapping using an F₂ population from ZL1929-4 × Towada, a QTL for cold tolerance was detected on the long arm of chromosome 7. The QTL explained 9 and 21% of the phenotypic variances in the F₂ and F₃ generations, respectively. Recombinant plants were screened for two flanking markers, RM182 and RM1132, in an F₂ population with 2,810 plants. Two-step substitution mapping suggested that the QTL was located in a 92-kb interval between markers RI02905 and RM21862. This interval was present in BAC clone AP003804. We designated the QTL as qCTB7 (quantitative trait locus for cold tolerance at the booting stage on chromosome 7), and identified 12 putative candidate genes.     

Modeling the effects of nitrogen deposition on carbon budget in two temperate forests

The NO_x input terrestrial ecosystems are increasing significantly induced by human activities, yet the understanding of the responses of carbon cycle to nitrogen deposition is still poor. The northern temperate forest ecosystems have seen the greatest changes in nitrogen inputs from atmosphere. It is necessary for us to understand how the carbon cycle would change under the nitrogen addition in temperate forests, as an important carbon sink. In this study, we present a biogeochemical process model, CEVSA2, and use this model to elucidate the key processes that may strongly influence the carbon budget response to anthropogenic nitrogen addition. The CEVSA2 model has included the effect of nitrogen on photosynthesis, carbon allocation, soil organic matter decomposition, etc. Our simulations show nitrogen addition stimulates the photosynthesis, net carbon sequestration, carbon accumulation in vegetation and soil, by contrary, the low level of nitrogen addition decreases the heterotrophical and total respiration. The long-term chronic nitrogen addition experiments show that the low and high level nitrogen addition would reduce the carbon sequestration and accumulation. The model failed to simulate the effect of nitrogen addition on plant mortality, the de-coupling of nitrogen and photosynthesis when nitrogen saturates. In addition, the responses of soil respiration to nitrogen deposition involve so many complex biochemical processes; however, we have little knowledge about them. Sequentially, there is large uncertainty of model simulation on the effect of nitrogen deposition on soil respiration. With increasing rates of anthropogenic nitrogen deposition, there is a strong need to understand links between nitrogen inputs and carbon cycle.     

Eco1 is important for DNA damage repair in S. cerevisiae

The cohesin network has an essential role in chromosome segregation, but also plays a role in DNA damage repair. Eco1 is an acetyltransferase that targets subunits of the cohesin complex and is involved in both the chromosome segregation and DNA damage repair roles of the network. Using budding yeast as a model system, we find that mutations in Eco1, including a genocopy of a human Roberts syndrome allele, do not cause gross defects in chromosome cohesion. We examined how mitotic and meiotic DNA damage repair is affected by mutations in Eco1. Strains containing mutations in Eco1 are sensitive to DNA damaging agents that cause double-strand breaks, such as X-rays and bleomycin. While meiotic crossing over is relatively unaffected in strains containing the Roberts mutation, reciprocal mitotic crossovers occur with extremely low frequency in this mutant background. Our results suggest that Eco1 promotes the reciprocal exchange of chromosome arms and maintenance of heterozygosity during mitosis.Key words: cohesin, recombination, double-strand break, acetyltransferase, Roberts syndrome     

不明原因男性不育人群中睾丸特异性乳酸脱氢酶基因突变的发现及其意义

为了研究睾丸特异性乳酸脱氢酶,即乳酸脱氢酶C4(LDH-C4)基因突变在男性不育发病中的作用,利用LDH-C4特异性底物对100名不明原因男性不育症患者的精子LDH-C4进行活性显色,用变性高效液相色谱(DHPLC)技术对LDH-C4活性低下的患者进行LDHC基因PCR产物的突变筛查,对DHPLC峰形异常的PCR产物进行序列测定.筛选到一组精子LDH-C4活性明显下降的患者,其中1名患者的LDHC基因PCR产物在DHPLC中呈异常洗脱峰.对这一PCR产物进行序列测定,发现患者LDHC基因第5外显子的115位碱基发生了T→A的杂合改变(GenBank登录号GU479375),该突变使LDHC基因的178位密码子由原来的TTG(编码亮氨酸)变为TAG(终止密码子),形成截短的C亚基.T克隆-测序进一步证实了该无义突变的杂合状态.这是在人类LDHC基因上发现的第一个突变,提示LDHC基因突变可能是男性不育发病的原因之一.     

棉花咖啡酸-O-甲基转移酶基因的原核表达及蛋白纯化鉴定

为获得大量高纯度的GhCOMT2蛋白以便研究其功能和性质,以pMD18-GhCOMT2质粒为模板,PCR扩增GhCOMT2基因的cDNA编码区,构建原核表达载体pET-28a-GhCOMT2,经酶切鉴定并测序后转化到大肠杆菌BL21 (DE3)中进行诱导表达,并采用Western blotting方法鉴定表达产物.结果表明:在大肠杆菌BL21(DE3)菌株中成功表达了与标签蛋白融合的GhCOMT2蛋白,大小约为40.062 kD,浓度为0.62 mg/mL.重组蛋白的最佳诱导条件为:0.2 mmol/L IPTG在16℃诱导12 h.重组蛋白以可溶形式高效表达,用蛋白标签亲和层析柱(His TrapTM HP)获得纯化重组蛋白,Western blotting分析表明其能与His多克隆抗体起特异性反应.     

棉花4-香豆酸辅酶A连接酶基因克隆及原核表达

本研究从棉花中克隆了一个4CL基因,命名为Gh4CL2(GenBank登录号为FJ848870)。研究结果表明:Gh4CL2基因cDNA全长2332bp,具有一个1725bp的开放阅读框,5′非编码区为64bp,3′非编码区为543bp,编码574个氨基酸,预测分子量约为62.106kD,等电点为5.94。氨基酸同源性分析发现,Gh4CL2与来自白杨、大豆和紫草的4CL一致性较高。进一步研究Gh4CL2基因的功能,构建了该基因的原核表达载体pET-28a-4CL2,经酶切鉴定后转化到大肠杆菌BL21(DE3)中。SDS-PAGE分析表明,最佳诱导表达条件为0.5mmol/LIPTG在37℃下诱导4h,重组蛋白主要以包涵体形式出现。     

基于多尺度遥感影像分割方法的湿地生态廊道设计

从生物多样性保护的角度,采用多尺度遥感影像分割方法中的人为干扰度模型,计算分割阈值确定湿地生态廊道的宽度,并结合聚类分析法分类的8种人为干扰类型,对建三江地区廊道结构设计进行了研究。结果表明,廊道分割阈值设为20%,非湿地背景噪声为9.43%,湿地生态廊道最佳宽度为1298m。8种人为干扰度中聚类c1、c2、c3和c4类型是受人为干扰较弱的区域,主要分布在浓江、乌苏里江、三江和洪河保护区原始生态环境区域,将其分别设定为核心区、实验区、边缘区、缓冲区4种类型。廊道核心区中的沼泽类型占75%,总体精度高达93.7%,实验区沼泽占72.2%,精度达到75.8%,边缘与缓冲区起到边缘护栏的作用,缓冲区宽度为945m,本研究为湿地生态廊道的建设与修复提供了可靠、科学的参考依据。     

采笋对大熊猫主食竹八月竹竹笋生长的影响

采笋是大熊猫栖息地内一种持续的人类干扰活动。为了评估采笋对大熊猫主食竹竹笋生长发育的影响,2009年7—11月,在四川省洪雅县瓦屋山镇设置实验样方,对比研究了早期采笋、中期采笋、晚期采笋、一直采笋和不采笋5种采笋方式对大熊猫主食竹八月竹竹笋生长和发育的影响。结果表明:1)早期采笋、中期采笋和一直采笋样方的发笋量显著高于不采笋样方(P0.05),其发笋量分别是不采笋的1.53倍,1.57倍和1.62倍;晚期采笋样方发笋量与不采笋样方差异不显著(P0.05)。2)早期采笋使八月竹在发笋早期和发笋中期的发笋量增加,中期采笋仅使八月竹在发笋中期的发笋量增加,而一直采笋使八月竹在整个发笋期的各个阶段的发笋量均显著增加(P0.05)。3)一直采笋样方的幼竹数量、株高均显著低于其他采笋样方,其基径只显著低于不采笋样方(P0.05);早期、中期、晚期采笋和不采笋样方相互间仅幼竹株高差异显著(P0.05)。4)早期、中期和晚期采笋与不采笋样方的发笋总量、退笋总量和采笋总量相互之间均呈显著正相关(P0.05)。建议仅在八月竹发笋中期采笋,同时政府应加大对大熊猫栖息地周边社区经济发展的引导和扶持。     

流动沙丘区和固定沙丘区丘间低地植物种间关联关系

丘间低地植物种间关联研究对揭示植物对生境变化的响应及植被演替机制具有重要意义。通过χ2检验和AC关联系数对科尔沁沙地22个不同面积的流动沙丘区和固定沙丘区丘间低地植物种间关联进行了比较分析。结果表明:在固定沙丘区,当丘间低地面积<0.5hm2时,|AC|≤0.3的种对数占所有种对数的百分比和|AC|≥0.7的种对数占所有种对数的百分比略大于流动沙丘区,当丘间低地面积>0.5hm2时,|AC|≤0.3的种对数占所有种对数的百分比显著小于流动沙丘区,|AC|≥0.7的种对数占所有种对数的百分比显著大于流动沙丘区;当面积<0.5hm2时,沙丘固定未引起丘间低地植物种间关系发生明显变化;当面积>0.5hm2时,沙丘固定导致丘间低地植物种间关系更加紧密。     

蕙兰根内可培养细菌的物种多样性

以MS基本培养基添加蕙兰菌根浸出液制成的培养基进行分离培养的方法,从野生蕙兰（Cymbidium faberi）根部首次分离到内生细菌。经过分离纯化培养获得纯菌株27株。经过16S rDNA基因序列测序,并与GenBank数据比对,其相似性均在98%以上,分析鉴定结果表明,存活的22株菌可分为8属14种。分别隶属于伯克氏菌属（Burkholderia）、假单胞菌属（Pseudomonas）、芽胞杆菌属（Bacillus）、Leifsonia属、贪食菌属（Variovorax）、欧文氏菌属（Erwinia）、Duganella属和不动杆菌属（Acinetobacter）。对这些菌株进行分离培养及鉴定有助于理解兰花与微生物之间的相互作用关系,为开发利用这些微生物开辟新的思路。