核糖体RNA基因在海洋动物分子系统学中的应用

随着分子遗传标记在分子系统学中应用的发展,核糖体RNA基因因其特殊的组成结构和演化方式被广泛应用于海洋动物分子系统学研究中.本文系统地介绍了真核生物核糖体RNA基因的组成和性质及其近年来在海洋动物系统进化、分类和种质鉴定、遗传多样性研究等方面研究中的应用.从中可以看出,核糖体RNA基因已经广泛应用于海洋动物的分子系统学研究中,但主要利用的是核糖体RNA基因的一级结构.图2参36     

粗山羊草间杂交及抗条锈病新基因遗传分析和分子标记

粗山羊草是小麦野生近缘属种,是D基因组的供体,蕴含大量的抗病资源,是进行小麦遗传改良的重要资源.选取条锈病免疫材料Y206和高度感病材料Y121杂交后代进行遗传分析和抗病性鉴定.从粗山羊草[Aegilops tauschii(Coss.)Schmal]Y206中鉴定出1个显性抗小麦条锈病基因,暂定名为YrY206.并利用SSR分子标记对该抗病基因标记定位,应用分离群体分组法(Bulked segregant analysis,BSA)筛选到Wmc11a、Xgwm71c、Xgwm161和xgwm183标记,与该基因之间的遗传距离分别为4.0 cM、3.3 cM、1.5 cM和9.3 cM.根据连锁标记所在小麦微卫星图谱的位置,YrY206被定位在3DS染色体上,可能是一个新的抗小麦条锈病基因.图2表2参22     

微卫星标记技术在淡水腹足类种群遗传学研究中的应用

综述了淡水腹足类微卫星位点的获取方法、微卫星重复序列的特点以及微卫星标记技术在其种群遗传学研究中的应用,着重分析了微卫星多态性在淡水腹足类种群遗传多样性、种群遗传结构、遗传分化、交配系统、种群关系研究中的应用.淡水腹足类普遍表现为杂合体缺失,种群遗传多样性降低.淡水腹足类近交和自体受精现象比较频繁以及地理隔离导致种群间遗传分化.利用微卫星标记研究遗传杂合度,结合群体近交系数和基因分化系数在一定程度上反映了淡水腹足类的交配系统.根据不同种群的遗传距离和相似性进行聚类分析在判别种群间的亲缘关系上起到了很大作用.针对淡水腹足类自身的特性,微卫星分子标记技术在淡水腹足类入侵生物学、分类阶元,寄生虫的分子流行病学等研究领域有广泛的应用前景.     

水稻Wx基因PCR-Acc Ⅰ标记与稻米AC的关系及其辅助育种效果

利用PCR-AccⅠ分子标记分析了105个水稻品种的Wx基因型,结果表明,用该标记检测的Wx基因型与该品种的稻米直链淀粉含量有较好的对应关系,利用PCR-AccⅠ标记可以鉴别籼稻品种直链淀粉含量的高或低;同时,对2个杂交组合F2分离群体的分析表明,PCR-AccI标记与稻米直链淀粉含量是紧密连锁、共分离的.另一方面,以直链淀粉含量中等的优质籼稻保持系D香1B为优质Wx基因供体,运用PCR-AccⅠ分子标记辅助选择,对综合性状优良、配合力高,但直链淀粉含量过高、品质欠佳的籼稻保持系G46B进行品质改良,结果在BC3F2代成功获得了直链淀粉含量中等的纯合TT基因型目标植株,表明PCR-AccⅠ标记用于优质Wx基因的分子标记辅助选择育种是有效的,因而该标记对改良稻米品质有重要作用.图2表3参13     

岩生植物金发草遗传多样性的ISSR和AFLP比较研究

禾本科岩生植物金发草〔Pogonatherum paniceum(Lam.)Hack.〕具有开发为新型国产草坪草种的潜力.用IS-SR与AFLP分子标记,分析了四川、重庆、云南及广西等地22个金发草种群的遗传多样性.用14个ISSR引物、10对AFLP引物分别扩增出239、485条谱带,多态位点百分率分别为94.98%、90.93%.两种标记均揭示出金发草种群具有丰富的遗传多样水平.由于标记原理不同,两种标记的结果呈现出微小的差别.相对而言,基于ISSR标记分析的遗传多样性参数高于AFLP标记得到的遗传多样性参数,这可能是由于金发草种群间存在着大量的微卫星变异所致.两种标记基于Nei遗传距离的聚类分析结果存在着一定的差异,但用Mantel测试对两种方法检测的遗传一致度进行相关性分析表明,它们之间存在着显著的相关性(r=0.3146,P=0.0150),用SPSS11.5软件进一步分析两种标记遗传一致度的相关性,Pearson相关系数为0.315(在0.01水平上),两种标记的聚类分析都揭示出金发草的遗传变异呈现出一定的地域分布规律.ISSR标记与AFLP标记均能应用于金发草种群的遗传多样性研究.图1表2参16     

屋顶绿化条件下四种草坪草耗水特性及土壤水分阈值

本文以4种草坪草——高羊茅、黑麦草、狗牙根、中华结缕草为试验材料,在屋顶环境下应用小型蒸散仪和便携式光合测定系统,对4种草坪草不同土壤水分梯度下的光合生理参数的光响应进程进行测定.结果表明,暖季型草坪草的光合速率和水分利用效率均高于冷季型草坪草;4种草坪草在屋顶环境下最佳土壤水分阈值为:高羊茅71.50%—87.50%、黑麦草59.70%—70.30%、狗牙根56.00%—82.60%、中华结缕草71.50%—89.00%.制定了适宜的灌水方案,在草坪草生长季内,黑麦草、高羊茅两种冷季型草坪草分别需补充灌水次数为33次和37次,每次19.29 mm和16.64 mm;狗牙根和中华结缕草两种暖季型草坪分别需补充灌溉25次和27次,每次20.75 mm和18.20 mm.本研究以期为今后草坪草的节水灌溉与水分的量化管理提供参考依据.     

分子微生物生态学技术在中国油田开发中的应用

分子微生物生态学技术近年来在研究油藏环境中微生物群落和生态系统等方面得到了广泛的应用,已经成为指导微生物采油技术开发和应用的一个不可或缺的工具.本文较系统地总结了我国利用分子微生物生态学方法研究油藏微生物群落结构的进展,其中包括大庆油田和西北部油田的16SrRNA基因序列分析,胜利油田16SrDNA的荧光标记末端多形态限制性片断分析技术(T-RFLP),以及吉林油田应用的DNA基因跟踪检测技术,并指出了存在问题和研究方向.图1参18     

五氯酚暴露诱导斑马鱼胚胎细胞凋亡相关基因的变化及caspase-2基因克隆和系统进化分析

应用基因芯片技术研究发现,暴露于五氯酚的斑马鱼胚胎中有14个涉及细胞凋亡行为的基因表达发生了显著改变.利用生物信息学方法构建这些功能基因系统进化树,分析与环境毒物诱导的细胞凋亡行为密切相关的caspase-2基因和其他基因之间的同源关系.斑马鱼caspase-2基因由10个外显子和9个内含子组成,cDNA长1308bp,含一个开放阅读框(ORF),编码435个氨基酸.6种脊椎动物Caspase-2氨基酸序列的保守性及系统进化分析结果表明,在特定功能区结构域中氨基酸序列表现出较高的同源性,Caspase-2在进化上高度保守.利用RT-PCR技术,斑马鱼caspase-2基因cDNA被克隆并确认.研究结果表明斑马鱼caspase-2基因是一个研究细胞凋亡行为的模型分子,可为环境化合物的分子毒性评价及其机制研究提供一种分子标记。     

盐胁迫对3种冷季型草坪草生长的影响

冷季型草坪草在不同盐离子含量的胁迫下生长,其各项生理指标都会发生相应的变化,这些变化可以反应出不同草坪草抗逆性的强弱.本实验所用的品种为早熟禾P.Pratensis L.、黑麦草L.Perenne L.、高羊茅F.Arundinacea Schreb..3种草均为冷季型草坪草.将氯化钠溶液设为5个处理,其质量分数分别为0.1%、0.2%、0.3%、0.6%、0.9%和一个对照,每个处理3次重复.将处理后3种草坪草的种子,播于营养钵中,实验室内培养.每日浇灌相对应的盐溶液,浇灌量为约2/3的溶液流出.待幼苗长到5～7 cm时,进行测定.在盐胁迫条件下,通过对黑麦草、高羊茅、早熟禾的发芽率、POD酶活性、质膜相对电导率等生理指标测定的研究,结果表明黑麦草抗盐性较强,早熟禾的耐盐性较弱.为在盐碱地上种植草坪提供了参考依据.     

基因组编辑技术的原理及应用

阐明基因功能和改良生物现状是生物学重点研究内容之一.近年来利用的基因打靶和转基因技术存在效率低、耗时长、易引起人们的安全性疑虑等问题.新近发现的以多种新型高效的DNA靶向内切酶为基础而建立的基因组编辑技术,主要包括ZFN、TALEN和CRISPR/Cas三种.本文首先综述这3种技术的原理,即均基于"DNA断裂/DNA损伤修复"来实现编辑功能,可以在不同物种中对目标基因进行定点敲除、单核苷酸或多核苷酸片段的置换、添加等基因组靶向修饰,具有简单、快速、高效、准确等优点.然后比较3种技术在构成、靶点识别模式、编辑特点、技术难度以及脱靶效应等方面的异同;归纳该技术在作物遗传育种、家畜改良以及基因治疗等方面的应用现状和前景,并指出该技术尚存在的脱靶、构建等问题以及解决的途径,重点突出ZFN、TALEN和CRISPR/Cas在模式植物以及粮食作物中的应用.最后提出建立基于病毒载体的基因组无痕编辑技术平台,将使基因组编辑用于植物遗传改良更为简约和安全.     

锌、镉复合污染对重金属蓄集植物黑麦草养分吸收及锌、镉积累的影响

采用根袋盆栽试验研究了锌、镉单一及复合污染对重金属蓄集植物黑麦草(Lolium perenne L．)生长、养分吸收及锌、镉积累的影响．结果表明,锌、镉单一及复合污染对黑麦草生物量存在不同程度的抑制作用．锌、镉单一及复合污染均明显降低了黑麦草植株N、P、K含量,以锌、镉复合污染对植株养分吸收的抑制作用最大．锌、镉污染共存下,黑麦草对锌、镉的吸收为协同效应．黑麦草吸收的锌、镉主要集中在地上部,以锌、镉复合污染植株对锌、镉的蓄集量最大,其地上部锌、镉含量分别达到3108．72、73．97 mg·kg-1．     

Inbreeding Depression in the Speke's Gazelle Captive Breeding Program

Abstract: The Speke's gazelle ( Gazella spekei ) captive breeding program has been presented as one of the few examples of selection reducing the genetic load of a population and as a potential model for the captive breeding of endangered species founded from a small number of individuals. In this breeding program, three generations of mate selection apparently increased the viability of inbred individuals. We reanalyzed the Speke's gazelle studbook and examined potential causes for the reduction of inbreeding depression. Our analysis indicates that the decrease in inbreeding depression is not consistent with any model of genetic improvement in the herd. Instead, we found that the effect of inbreeding decreased from severe to moderate during the first generation of inbreeding, and that this change is responsible for almost all of the decline in inbreeding depression observed during the breeding program. This eliminates selection as a potential explanation for the decrease in inbreeding depression and suggests that inbreeding depression may be more sensitive to environmental influences than is usually thought.     

Accumulation and phytoavailability of benzo[a]pyrene in an acid sandy soil

Effects of benzo[a]pyrene (B[a]P) on ryegrass (Lolium perenne L.) growth, plant accumulation and dissipiation of B[a]P in a red sandy soil (Hapli-Udic Argosol) were studied in a pot experiment. The plants were grown for 61 days in soil spiked with B[a]P at 0, 12.5, 25 and 50 mg kg⁻¹. Control pots without plants were also set up. Soil extractable B[a]P, plant shoot and root biomass, and concentrations of B[a]P in plant shoots and roots were determined. Ryegrass biomass was increased by addition of B[a]P and root B[a]P concentrations were significantly correlated with B[a]P application rate, but no such correlation was found for shoot B[a]P concentrations. This indicates that B[a]P enhanced the growth of the ryegrass. The extractable B[a]P concentration in the planted soil was significantly lower than that in the unplanted control soil at the rate of 50 mg B[a]P kg⁻¹. This indicates that ryegrass may help to dissipate B[a]P in soil at concentrations over 50 mg kg⁻¹ soil although the mechanism for this is not understood.     

Use of degenerate primers in rapid generation of microsatellite markers in Panicum maximum

Guineagrass (Panicum maximum Jacq.) is an important forage grass of tropical and semi-tropical regions, largely apomictic and predominantly exist in tetraploid form. For molecular breeding work, it is prerequisite to develop and design molecular markers for characterization of genotypes, development of linkage map and marker assisted selection. Hence, it is an important researchable issue to develop molecular markers in those crops where such information is scanty. Among many molecular markers, microsatellites or simple sequence repeat (SSR) markers are preferred markers in plant breeding. Degenerate primers bearing simple sequence repeat as anchor motifs can be utilized in rapid development of SSR markers; however selection of suitable degenerate primers is a prerequisite for such procedure so that SSR enriched genomic library can be made rapidly. In the present study seven degenerated primers namely KKVRVRV(AG)10, KKVRVRV(GGT)5, KKVRVRV(CT)10, KKVRVRV(AAT)6, KKVRVRV(GTG)6, KKVRVRV(GACA)5, and KKVRVRV(CAA)6 were used in amplification of Panicum maximum genomic DNA. Primers with repeat motifs (GGT)5 and (AAT)6 have not reacted whereas (AG)10, (GACA)5 and (CAA)6 highly informative as they have generated many DNA fragments ranging from 250 to 1600 bps as revealed from the results obtained with restriction digestion of recombinant plasmids. Primer with (CT)10 anchor repeat, amplified fragments of high molecular weight where as (GTG)6 primer generated only six bands with low concentration indicating less suitability of these primerin SSR markers development in P maximum.     

4种改良剂对土壤-黑麦草系统中镉行为的影响

以黑麦草（Lolium perenne L.）作为修复植物,开展了石灰、磷灰石、木炭、猪粪4种改良剂对重金属Cd污染土壤的田间原位修复试验研究。结果表明：改良剂提高了土壤pH值并促进土壤Cd从生物可利用性高的形态向迟效态转化,降低对生物和环境的直接毒害作用,促进黑麦草的生长。以黑麦草生物量和Cd富集量的增加作为评价指标,4种改良剂修复Cd污染土壤效果依次为：石灰〉磷灰石〉木炭〉猪粪。石灰处理中黑麦草Cd富集量为13.11 mg,与其他处理均呈现显著差异。污染土壤重金属Cd的化学形态与黑麦草对Cd的吸收密切相关。黑麦草地上部分Cd质量分数,与土壤B-2态Cd呈极显著正相关关系,相关系数为0.730。黑麦草地下部分Cd质量分数,与B-2、B-3态Cd均呈极显著正相关关系,相关系数分别为0.756、0.786;黑麦草地上部分和地下部分Cd质量分数均与B-4态Cd呈极显著负相关关系,相关系数分别为0.757和0.708。     

Genetic Variation for Morphological and Allozyme Variation in Relation to Population Size in Clarkia dudleyana, an Endemic Annual

Abstract: The maintenance of genetic variation within populations is expected to allow species to respond to evolutionary challenges such as selection and environmental stress. Larger populations are generally expected to maintain larger amounts of genetic variation. Although several studies have found a positive relationship between population size and levels of genetic variation for molecular markers such as allozymes, few comparisons have been made between molecular measures of variation and genetic variation that is likely to be ecologically important. Most ecologically important traits require quantitative genetic analyses. I examined the relationship between levels of genetic variation and population size for both allozymes and morphological traits in a California endemic annual plant, Clarkia dudleyana . Levels of genetic variation for allozymes did not show a significant positive relationship with population size. The level of genetic variance for all of the 18 morphological traits exhibited no significant relationship with population size. Further, allozyme heterozygosities were not related to levels of quantitative genetic variation. These results indicate that levels of allozyme variability do not predict levels of genetic variation for morphological traits in C. dudleyana , suggesting that molecular measures of variation, in general, differ from quantitative genetic measures. These results imply that conservation genetic studies should generally focus on aspects other than measuring levels of genetic variation found within populations.     

牧草轮作对奶牛场污水氮、磷的净化效果研究

以土壤为基质,选择象草（Pennisetum purpureum）和黑麦草（Lolium perenne）两种牧草作为植被模拟人工湿地净化系统,研究牧草全年轮作在间歇流人工湿地中对奶牛场高质量浓度污水的适应性及对污水氮、磷的净化效果。结果表明：象草、黑麦草在间歇性高质量浓度污水中生长良好,可作为畜牧场高质量浓度污水修复植物进行全年轮作,并适时的收割,去除污水中的氮、磷。在不同污水质量浓度处理的牧草净化系统中,水力停留时间为8 d的条件下,铵氮的去除率为75.9%~94.9%;硝态氮的去除率为65.4%~90.9%;总氮的去除率64.2%~90.1%;总磷的去除率在90%以上;象草和黑麦草对污水中氮、磷的吸收转化在系统对氮磷净化中的贡献率平均分别为氮25.12%和20.03%、磷31.20%和20.42%;各处理氮、磷的去除效果与污水的质量浓度呈负相关。     

无瓣海桑和互花米草群落土壤化感作用比较

为揭示不同植物群落土壤在无瓣海桑(Sonneratia apetala)控制互花米草(Spartina alterniflora)中的作用,利用化感生物测定的方法,选用黑麦草(Lolium perenne)为受体,对比分析红树林无瓣海桑群落、互花米草群落以及无瓣海桑+互花米草群落土壤水提液的化感作用大小。结果表明3,种植物群落土壤水提液对黑麦草具有化感作用,无瓣海桑群落土壤和无瓣海桑+互花米草群落土壤水提液对黑麦草根长和苗高的影响表现出低促高抑的现象;ρ为0.25 g·mL-1的土壤水提液对黑麦草种子发芽和幼苗生长影响不大,1.0 g·mL-1土壤水提液对黑麦草根长和苗高产生显著影响,加入活性碳之后能减弱水提液的化感作用。ρ为0.25或0.5 g·mL-1条件下3,种土壤浸提液对黑麦草发芽率的影响无显著差异;ρ为1.0 g·mL-1的3种水提液都显著抑制黑麦草根长,而无瓣海桑群落土壤水提液表现出更强的化感作用,ρ为1.0 g·mL-1的土壤水提液显著抑制黑麦草苗高,无瓣海桑群落土壤对苗高的抑制作用显著强于互花米草群落土壤(P<0.01)。以黑麦草为受体,无瓣海桑群落土壤化感作用强于互花米草群落土壤,通过根系分...     

硅对镉、锌、铅复合污染土壤中黑麦草生理生化性质的影响

采用正交试验设计L9(34)对黑麦草(Lolium perenne L.)进行温室盆栽试验,观测Cd、Zn、Pb复合污染条件下,硅(Si)对黑麦草生物量、叶绿素含量以及保护酶系统的影响.结果表明,Cd使黑麦草根系发育受阻导致生物量下降,中等水平的Zn和Pb有利于黑麦草生物量的积累;Cd、Zn、Pb复合污染造成黑麦草叶绿体结构的破坏,使叶绿素含量减少;低水平的Cd、Zn、Pb复合污染对黑麦草CAT和POD活性起到激发作用,但随着添加水平提高,两种酶活性受到抑制.Si可以促进黑麦草根系生物量的增加,有利于根系对水分和养分的吸收,保证地上部分的养分供给,使叶片生物量增加;硅化细胞的形成有利于黑麦草叶片对光能的吸收利用,Si也使叶绿素含量增加;Si对CAT和POD有显著激活作用,从而减轻重金属复合污染对黑麦草产生的伤害.     

Morphological and molecular variation within an ocean basin in wedge-tailed shearwaters (Puffinus pacificus)

Micro-evolutionary processes that underpin genetic and morphological variation in highly mobile pelagic vertebrates are virtually unknown. Previous findings preferentially invoke vicariant isolation due to large-scale physical barriers such as continental landmasses, followed by genetic drift. However increasingly, evidence for divergence by non-random processes (e.g. selection, plasticity) is being presented. Wedge-tailed shearwaters are wide-ranging seabirds with breeding colonies located such that they experience a variety of environmental pressures and conditions. Previous work on this species has provided evidence of inter-colony divergence of adult morphology and foraging modes, as well as chick developmental patterns, suggesting that reinforcement among colonies is possible. In order to evaluate the micro-evolutionary processes driving this observed variation, our study compared patterns of gene flow with morphological and environmental variation among four colonies of wedge-tailed shearwater breeding within the Indo-Pacific Ocean basin. Estimates of gene flow differed according to the genetic marker used; most likely, this is a function of different mutation rates. Nuclear introns suggest that gene flow among wedge-tailed shearwater breeding colonies within the Indo-Pacific Ocean basin is substantial, however microsatellite markers imply that gene flow is reduced. In general, levels of genetic divergence were relatively low and did not correlate with geographic distance, morphological distance or environmental differences (sea-surface temperature and chlorophyll a concentration) among colonies. We suggest that genetic drift alone is unlikely to be the major source of morphological variation seen in this species. Instead, we propose that non-random processes (selection, plasticity) underpin morphological diversity seen in this and possibly other seabird species.