Alien DNA introgression and wheat DNA rearrangements in a stable wheat line derived from the early generation of distant hybridization

In general, it requires five or more generations to develop a stable line by the traditional breeding method of cultivar hybrids in self-pollinated crops, such as wheat, barley, and rice. No doubt, the breeding period will be shortened and thus the breeding effi-ciency will be improved if the hybrids can stabilize in early generation. The method of haploid breeding, that is, obtaining stable pure lines (DH lines) of cultivar hybrids with the aid of anther culture and then chro-mosome doubling,…     

T-box genes coordinate regional rates of proliferation and regional specification during cardiogenesis

Mutations in T-box genes are the cause of several congenital diseases and are implicated in cancer. Tbx20-null mice exhibit severely hypoplastic hearts and express Tbx2, which is normally restricted to outflow tract and atrioventricular canal, throughout the heart. Tbx20 mutant hearts closely resemble those seen in mice overexpressing Tbx2 in myocardium, suggesting that upregulation of Tbx2 can largely account for the cardiac phenotype in Tbx20-null mice. We provide evidence that Tbx2 is a direct target for repression by Tbx20 in developing heart. We have also found that Tbx2 directly binds to the Nmyc1 promoter in developing heart, and can repress expression of the Nmyc1 promoter in transient transfection studies. Repression of Nmyc1 (N-myc) by aberrantly regulated Tbx2 can account in part for the observed cardiac hypoplasia in Tbx20 mutants. Nmyc1 is required for growth and development of multiple organs, including the heart, and overexpression of Nmyc1 is associated with childhood tumors. Despite its clinical relevance, the factors that regulate Nmyc1 expression during development are unknown. Our data present a paradigm by which T-box proteins regulate regional differences in Nmyc1 expression and proliferation to effect organ morphogenesis. We present a model whereby Tbx2 directly represses Nmyc1 in outflow tract and atrioventricular canal of the developing heart, resulting in relatively low proliferation. In chamber myocardium, Tbx20 represses Tbx2, preventing repression of Nmyc1 and resulting in relatively high proliferation. In addition to its role in regulating regional proliferation, we have found that Tbx20 regulates expression of a number of genes that specify regional identity within the heart, thereby coordinating these two important aspects of organ development.     

Semaphorin 3B is a 1,25-Dihydroxyvitamin D3-induced gene in osteoblasts that promotes osteoclastogenesis and induces osteopenia in mice

The vitamin D endocrine system is important for skeletal homeostasis. 1,25-Dihydroxyvitamin D(3) [1,25(OH)(2)D(3)] impacts bone indirectly by promoting intestinal absorption of calcium and phosphate and directly by acting on osteoblasts and osteoclasts. Despite the direct actions of 1,25(OH)(2)D(3) in bone, relatively little is known of the mechanisms or target genes that are regulated by 1,25(OH)(2)D(3) in skeletal cells. Here, we identify semaphorin 3B (SEMA3B) as a 1,25(OH)(2)D(3)-stimulated gene in osteoblastic cells. Northern analysis revealed strong induction of SEMA3B mRNA by 1,25(OH)(2)D(3) in MG-63, ST-2, MC3T3, and primary osteoblastic cells. Moreover, differentiation of these osteogenic cells enhanced SEMA3B gene expression. Biological effects of SEMA3B in the skeletal system have not been reported. Here, we show that osteoblast-derived SEMA3B alters global skeletal homeostasis in intact animals and osteoblast function in cell culture. Osteoblast-targeted expression of SEMA3B in mice resulted in reduced bone mineral density and aberrant trabecular structure compared with nontransgenic littermates. Histomorphometry studies indicated that this was likely due to increased osteoclast numbers and activity. Indeed, primary osteoblasts obtained from SEMA3B transgenic mice stimulated osteoclastogenesis to a greater extent than nontransgenic osteoblasts. This study establishes that SEMA3B is a 1,25(OH)(2)D(3)-induced gene in osteoblasts and that osteoblast-derived SEMA3B impacts skeletal biology in vitro and in vivo. Collectively, these studies support a putative role for SEMA3B as an osteoblast protein that regulates bone mass and skeletal homeostasis.     

东方蜜蜂微孢子虫侵染过程中nce-miR-34537和靶基因PIP5KI的表达谱

【目的】本研究旨在对前期鉴定到的nce-miR-34537进行表达和序列验证,预测nce-miR-34537的靶基因并明确其分子特性,进而检测nce-miR-34537及其靶基因在东方蜜蜂微孢子虫(Nosema ceranae)侵染意大利蜜蜂(Apis mellifera ligustica)工蜂过程的表达谱,为进一步探究nce-miR-34537调控东方蜜蜂微孢子虫侵染的功能和作用机制提供基础。【方法】通过Stem-loop-RT-PCR和Sanger测序验证nce-miR-34537的表达和序列。通过生物信息学软件预测nce-miR-34537的靶基因PIP5KI(I型磷脂酰肌醇4-磷酸-5-激酶基因)的理化性质等分子特性和保守基序,并构建基于氨基酸序列的系统进化树。采用RT-qPCR检测nce-miR-34537及其靶基因的表达谱。【结果】nce-miR-34537在东方蜜蜂微孢子虫孢子中真实存在和表达。nce-miR-34537共靶向PIP5KI等151个基因。PIP5KI蛋白的分子式为C₈₈₂H_{1 364}N₂₂₆     

湘西“蒿菜粑粑”原料植物鼠麴草总黄酮提取物体外抗氧化活性研究

为了解湘西特色食品“蒿菜粑粑”原料植物鼠麴草(Gnaphalium affine)总黄酮提取物体外抗氧化能力,采用DPPH、ABTS自由基清除实验,还原力实验和抑制β-胡萝卜素褪色实验等方法,测定鼠麴草总黄酮抗氧化活性。结果显示,鼠麴草总黄酮母液中总黄酮浓度为7.01 mg·mL–1mg/mL;总黄酮提取物对DPPH、ABTS自由基有较好的清除能力,其半数抑制浓度(IC50)分别为16.30 mg·L–1、30.16 mg·L–1,将胡萝卜素相对吸光度降为50%的时间延长至67.49 min,在还原能力、延缓胡萝卜素褪色和抑制脂质过氧化上也有较好效果。鼠麴草总黄酮提取物具有良好的体外抗氧化活性,可作为优质食用植物资源进一步开发与推广。     

利用短短小芽孢杆菌启动子和信号肽编码序列构建穿梭分泌表达载体

以短短小芽孢杆菌B15的总DNA为模板,利用PCR技术克隆到其细胞壁蛋白基因串联启动子和信号肽编码序列,测序分析后提交GenBank,登录号为AY956423。重新设计引物扩增该片段并在PCR产物两侧引入BamHⅠ和PstⅠ酶切位点,将PCR产物双酶切后克隆至穿梭载体pP43NMK的相应位点构建分泌表达载体pP15MK,插入片段置于该载体中mpd基因的上游,并使信号肽编码序列与去除了自身信号肽编码序列的mpd基因阅读框恰好融合。将pP15MK导入枯草杆菌构建表达菌株1A751(pP15MK),在短短小芽孢杆菌启动子和信号肽元件的带动下,mpd基因能够在表达菌株的对数生长期和稳定期持续性高效分泌表达,表达产物结合在细胞膜上;发酵液在48h酶活达到最高值7.79U/mL,是出发菌株邻单胞菌M6表达量的8.1倍。     

鉴定9个新的RHD基因mRNA可变剪接体

为了研究各种RHD基因mRNA可变剪接体的基因结构, 应用逆转录聚合酶链反应(RT-PCR)检测正常人脐血样本RHD mRNA, 对RHD cDNA进行TA克隆和序列分析, 对各可变剪接体的剪接位点进行DNA序列分析, 并将RHD mRNA进行表达序列标签(ESTs)分析。结果在28个阳性克隆中, 除全长RHD cDNA外, 共检测到12种(包括9种新的)RHD可变剪接体, 发现外显子遗漏、5′和3′剪接位点变异3种剪接形式, 涉及外显子2~9, 其中6种新的剪接体同时存在RHD和RHCE基因同源杂交现象。ESTs分析还检索到内含子保留形式的剪接体。研究表明, RHD基因mRNA存在复杂的可变剪接机制, 除已报道的剪接体外, 检测到9种新的RHD可变剪接体, 并发现了可变剪接和同源杂交并存现象。     

开花对两种杓兰光合作用和同化产物分配的影响

对同一生境下黄花杓兰（Cypripedium flavum）和西藏杓兰（Cypripedium tibeticum）开花株与非开花株的气体交换、光系统Ⅱ实际光化学效率（φPSⅡ）和^14C分配进行了测定,目的在于了解开花对杓兰光合作用以及同化产物分配的影响。结果表明这两种杓兰开花均能通过提高电子传递速率来提升植株的光合作用。相对于黄花杓兰,西藏杓兰开花显著提高植株的呼吸速率,以致其最大净光合速率（Amax）的提升不显著。两种杓兰的花和地下部分（包括根状茎和芽）在同化产物分配上存在竞争关系,因而开花减少了同化产物向植株地下部分的分配。     

西南岩溶地区石漠化综合治理研究

我国西南岩溶地区石漠化是人地矛盾作用下生态系统退化的过程或结果,石漠化不仅带来水土流失加剧、旱涝灾害频发、生物多样性降低等生态问题,还会带来贫困落后等一系列社会问题。20世纪80年代开始进行的石漠化治理工作已经成为我国西南岩溶地区生态文明建设的重要举措与必由之路,通过近40年来的石漠化治理使得我国西南岩溶地区石漠化问题恶化的趋势开始得到扭转。面向生态文明新时期的建设目标,石漠化脆弱区仍存在人地矛盾难以全面消除、治理成果可持续性亟待提升等问题,石漠化防治需要从多年来石漠化治理研究与实践探索积累中汲取有益于促进人地和谐、提升生态系统可持续性的治理经验与做法。因此,本文以石漠化治理典型区域调研资料与文献资料为数据基础,从石漠化治理目标、技术、措施和模式四个层面梳理石漠化治理经验与做法,归纳总结出石漠化综合治理四点配置原则:(1)以治理目标与预期周期为导向,确立石漠化治理采取的措施;(2)针对解决的关键生态退化问题,结合区域特征遴选石漠化治理技术;(3)针对石漠化退化阶段的差异,确定石漠化治理途径措施;(4)结合石漠化综合治理模式的共性特征,以小流域为基本单元,因地制宜的选择与构建石漠化综合治理模式。以期通过对实践经验的总结为石漠化治理过程中选择具备区位适宜性和机理明晰性的生态技术提供参考。     

厌氧菌Sunxiuqinia sp. SH-52外切型海藻酸裂解酶的异源表达及酶学特征

【背景】目前已报道的海藻酸分解菌多数为好氧菌,未见有关厌氧菌的报道。从分离的海藻酸分解菌中表征的海藻酸裂解酶大多为内切型海藻酸裂解酶,外切型较少。【目的】研究来自厌氧海藻酸分解菌的海藻酸裂解酶基因,表征新型海藻酸裂解酶并阐明其酶学性质,为海藻酸裂解酶的多样性和微生物降解海藻酸机制提供理论依据。【方法】对来自厌氧海藻酸分解菌Sunxiuqinia sp. SH-52的海藻酸裂解酶SHA-4编码基因进行克隆,分析基因序列,构建重组质粒PGEX-4T-1-SHA-4并在大肠杆菌中实现异源表达,经纯化后对其酶学性质及降解特征进行研究。【结果】该酶在28°C、用0.1 mmol/L IPTG (异丙基-β-D-硫代半乳糖苷)条件下诱导6 h达到最大表达量,纯化后酶的比活力达到21 U/mg。酶学性质分析表明SHA-4的最适温度为37°C;最适pH为7.5;对PolyMG (杂聚古罗糖醛酸-甘露糖醛酸嵌段)具有底物偏好性;Na+对该酶的活性具有抑制作用,而金属离子Cu~(2+)具有明显促进作用,使活性提高了约168%;SHA-4催化海藻酸的Km值为2.5 mg/mL,Vmax为8.7 mg/(mL·min);SHA-4为外切型海藻酸裂解酶,降解海藻酸终产物为单糖。【结论】异源表达了来自一株厌氧海藻酸分解菌Sunxiuqiniasp.SH-52的海藻酸裂解酶SHA-4,该酶是PL6家族中第一个对PolyMG有底物偏好性的外切型海藻酸裂解酶,而且活性较高,作为工具酶有很好的应用前景,为海藻酸降解机制的探索提供了新的线索。