柚类种质资源RAPD标记研究的引物筛选

利用 10 0个 10碱基随机引物 ,对柚类 4个品种酸柚、沙田柚、文旦柚和泰国柚进行了RAPD标记的引物筛选研究 ,结果为无扩增产物的引物 18个 ,在 1、 2、 3个和所有 4个样品中有扩增产物的引物数分别为 2 0、 13、 2 5和 2 4个 ;读取了 12个在所有 4个样品中都有扩增产物的引物的 RAPD带 ,计算了样品间 RAPD多态性位点的百分率为 60 .6% ;计算了样品间的相似系数和遗传距离 ,并对遗传距离进行了 UPGMA聚类分析 ,论证了利用所筛选出的引物对柚类进行 RAPD标记研究的可行性和可靠性     

怀地黄SRAP扩增体系的建立与引物的筛选

为建立适合怀地黄SRAP-PCR分子标记技术体系,通过单因子实验分别研究了DNA模板浓度、TaqDNA聚合酶浓度、Mg2+浓度、引物浓度以及dNTP浓度对怀地黄SRAP扩增反应的影响,确立了适合怀地黄SRAP最佳反应体系为:在25μL的反应体系中,模板DNA量20ng/25μL、2.5mmol/LMg2+、0.32μmol/L的上下游引物、0.30μmol/L的dNTP以及2.5UTaq酶,并利用确定的体系从88个引物组合中筛选出12对适合怀地黄SRAP-PCR反应的引物。     

甜瓜抗白粉病基因SRAP分子标记筛选

以感白粉病甜瓜A120和抗白粉病甜瓜A119为亲本构建F2代分离群体,采用BSA和SRAP相结合的方法筛选与甜瓜抗白粉病基因相连锁的分子标记.结果显示,294条SRAP引物中有6条引物在抗病与感病池间表现出多态性;对8个高抗和8个高感单株进行扩增,引物me46em51和me3em6分别在高感单株中扩增出210 bp和205 bp的多态性条带,而高抗单株无此扩增带,与抗病、感病池结果一致.采用JoinMap3.0软件进行连锁分析,两标记与抗白粉病基因的连锁距离分别为18.2 cM和23.4 cM,初步推测本实验中甜瓜白粉病抗性为隐性多基因控制.     

南药益智SRAP-PCR体系优化及引物筛选

为了建立适用于南药益智的SRAP-PCR体系,并筛选特异性引物用于研究不同地理居群的南药益智的遗传多样性,采集了海南不同地理居群的益智资源,利用植物基因组DNA提取试剂盒提取益智基因组DNA,并检测其纯度和浓度。采用正交试验对SRAP-PCR体系进行优化,最终建立了最优反应体系（总体积为25 μL）：Taq酶0.5 U,dNTPs 4 mmol/L,Mg2+ 2 mmol/L,引物各2 μmol/L,DNA模板10 ng,10×PCR buffer 2.5 μL。再利用最优反应体系对144对引物进行筛选,共获得7对特异性引物,其扩增片段的大小均在100~2 000 bp以内且分布较均匀,多态性条带比率皆达85%以上。该研究结果为南药益智遗传多样性的研究奠定了基础。     

虎杖种质资源的分子标记研究

本文应用RAPD、ISSR和SRAP标记对26份虎杖种质资源遗传多样性进行检测.在22个引物中有17个引物(77.3%)扩增产物具多态性,多态性水平相对较高.22个引物共得到98条扩增DNA片段,其中90.8%具有多态性.每个多态性引物平均可扩增出5.24个多态性片段.聚类分析表明,利用BAPD、ISSR和SRAP技术相结合可将全部供试材料区分开,26份材料在Gs值0.54水平上全部聚为一类,以所有材料间的平均遗传相似遗传系数0.71为阈值,将其分为11类.虎杖种质资源在分子水平上确实存在较大遗传差异,RAPD、ISSR和SRAP标记可作为构建虎杖DNA指纹图谱的有效工具.     

北极狐SRAP分子标记的多态性初步研究

相关序列多态性（SRAP）是近年来发展起来的一种新型分子标记技术,具有稳定、简便、中等产率、高共显性等优点。实验利用北极狐的基因组为模版,首次把SRAP方法引入到哺乳动物中进行分析,对北极狐SRAP反应条件进行了优化,确立了北极狐SRAP-PCR的反应条件是：94℃预变性5min,94℃1min,350C1min,72℃2min,5个循环;940C变性1min,94℃1min,55℃1min（根据不同的引物设定）,72℃1min,35个循环,72℃延伸10min,最后产物用6％的非变性的聚丙烯酰胺凝胶分离,得到的结果清晰、稳定、多态性高,条带可以用在后续对北极狐的遗传多样的分析和品种鉴定上。     

葡萄SRAP反应体系优化及引物筛选

本试验利用L16(45)正交试验设计探寻葡萄SRAP-PCR反应体系中的关键因子,同时结合单因素试验简单、快捷的特点逐个对PCR反应体系的主要成分进行优化.充分利用两种方法的优点并降低试验工作量取得了较好的效果,建立了适于葡萄的SRAP反应体系.结果表明Mg2+浓度为影响葡萄SRAP-PCR反应的关键因素:优化的20 μL SRAP-PCR反应体系中各组分的最适含量为:10×Buffer 2.0μL,Mg2+2.5mmol/L,dNTPs 0.3 mmol/L,引物0.4 μmol/L,DNA聚合酶1.0 U,模板DNA 1.0 ng/L.利用SRAP反应体系,从100对SRAP引物组合中筛选出扩增稳定,条带清晰,多态性好的引物19对.本研究建立的适于葡萄SRAP-PCR扩增的反应体系,将为葡萄种质遗传多样性评价、基因组分析、指纹图谱构建,分子标记辅助育种和遗传改良研究提供基础.     

基于SRAP标记的山药种质资源遗传多样性分析

目的:研究我国山药种质资源遗传多样性,为合理利用资源和开展选育种工作提供理论依据。方法:以国内94份山药种质资源为材料,采用SRAP标记并通过NTSYS2.10软件进行SHAN聚类分析、PROJECTION主成分分析;利用POPGENE软件估算遗传多样性参数。结果:从49对SRAP引物中筛选出30对能产生稳定清晰可辨的扩增产物的引物,共扩增出754条DNA带,其中多态性条带616条,占总条带的81.7%。聚类结果表明:当遗传相似系数(GS)为0.822时,可将94份资源分为5类:第Ⅰ类20份、第Ⅱ类43份、第Ⅲ类7份、第Ⅳ类3份、第Ⅴ类21份。第Ⅰ、Ⅱ、Ⅲ、Ⅴ类分别为薯蓣、褐苞薯蓣、山薯和参薯。主成分分析结果显示:第一与第二主成分可解释88.34%(82.10%和6.24%)的遗传总变异。遗传多样性参数分析表明:比较5个遗传多样性参数值,5个群体的遗传多样性水平表现为Ⅰ>Ⅴ>Ⅲ>Ⅱ>Ⅳ,第Ⅰ类(薯蓣)遗传多样性水平高;山药遗传群体间遗传分化系数为51.88%,大部分差异存在于群体之间,群体间遗传分化高。结论:山药种质资源丰富且群体遗传分化高,有利于山药新品种的选育。SRAP标记可有效应用于山药种质资源的鉴别和遗传多样性分析。通过DNA指纹鉴定技术鉴别山药品种具有重要性与紧迫性。     

SRAP分子标记分析西瓜遗传多态性

目的:探讨西瓜遗传多样性和遗传基础。方法:采用SRAP分子标记对西瓜品种D1、D2、D3、H1、H2、H3、M1、M2、M3、m1、m2、m3的多态性进行了分析。结果:每对引物组合产生13~25对比较清晰的扩增带.8对引物组合共产生131条扩增带。平均每对引物组合产生16.375条。8对引物组合共产生多态性带37条,每对引物组合产生3~7条,平均4.625条。每对引物组合产生的多态性带的比例为16.666%~38.464%,平均为28.675%。另外,对银染过程进行了优化。结论:SRAP标记多态性还是较高的,可以适于分析西瓜等遗传差异小的作物。     

草鱼种质相关SRAP及SCAR的分子标记

采用相关序列扩增多态性(Sequence-realted Amplified Polymorphism, SRAP)技术分析野生草鱼和家养草鱼,筛选与草鱼种质退化相关的分子遗传标记.共进行88对引物组合的检测, 产生标记数目共计905个.依据标记在群体中出现的频率和变化规律,共筛选出2 1个可能与种质相关的特异性标记,对这些特异性标记进行测序并将测序结果进行BLAST分析 .发现测得片段中有8个片段在GenBank中找到同源性较高的序列,而其他片段与数据库中序列的相似性较低.根据序列信息分别设计了3对引物.用这3对引物分别对草鱼三个群体进行 PCR扩增,分别产生了SCAR1(308 bp)、SCAR2(66 bp)、SCAR3(114 bp)3个扩增带.采用大样本对这3个标记进行验证,发现其中SCAR1在家养群体中呈现阳性,在野生群体中为阴性,可区分出这两种群体.以SCAR3为引物在174条家养群体中得到目的片段,在26个家养群体没有扩增出条带,分布频率为87%;在100个野生群体中有6个个体检测到该条带,分布频率为6%.以SCAR2为引物在野生群体中完全扩增出目的条带,淡水中心群体中有7条扩增到条带,前洲群体中没有扩增出条带,标记在家养种群中的分布频率为96.50%.因此SCAR1可作为草鱼家养群体的一个重要的分子遗传特征指标,为进一步进行分子标记辅助育种奠定了基础 [动物学报 54(3):475-481,2008].     

Effects of seismic stress on the vegetative growth of Glycine max (L.) Merr. cv. Wells II

Abstract. Vegetative plants of soybean [ Glycine max (L.) Merr. cv. Wells II] grown in a greenhouse and agitated periodically on a gyratory shaker had shorter stems, less leaf area, and lower leaf and plant dry weight than did undisturbed greenhouse-grown (GG) plants after 16 d of treatment. Outdoor-grown (OG) plants, which were subjected to additional environmental stresses including ultraviolet radiation, wind loading, and uncontrolled temperature and humidity fluctuations, were smaller and had less dry weight than GG controls, but growth was not inhibited further by gyratory shaking. Periodic shaking of GG soybeans resulted in the same plant and leaf dry weight as for OG soybeans. Response of GG plants to mechanical stress depended on light intensity, with minimum growth reduction occurring under full light (FL) level, and maximum growth reduction occurring under lower light levels (24–45% FL). Reduction in dry weight gain due to mechanical stress corresponded to a decrease in relative growth rate (RGR). Decreases in net assimilation rate and leaf area ratio contributed equally to the lower RGR of shaken plants, indicating that seismic stress inhibits dry weight accumulation by decreasing both the photosynthetic efficiency and the assimilatory surface of soybean.     

Effect of sulfur-containing agrochemicals on growth,yield, and protein content of soybeans (Glycine max (L.) Merr)

In this study, effect of different forms of sulfur-containing agrochemicals on growth, yield, and protein content of soybean grains have been evaluated. Three forms were used, such as powdery, solute, and pasty, in which elemental sulfur is contained in a nanostructured state. Plants treated with powdered and solute sulfur-containing agrochemicals had the highest growth and grain yield values, and the effect of applying pasty sulfur-containing agrochemicals did not differ from the control, in which there was low yield on all variants. The use of powdered and solute sulfur-containing agrochemicals increased all protein fractions in soybeans. The results show that the use of powdered and solute sulfur-containing agrochemicals is necessary to boost the yield of soy and increase the supply of proteins in the grains. A key factor in the availability of sulfur for soybean plants is the conversion of sulfur to a nanodisperse state. This study provides relevant information about sulfur-containing agrochemicals, which can promote higher seed yields and increase the content of protein in soybeans.     

Mapping QTLs for tissue culture response in soybean (Glycine max (L.) Merr.)

Hempseed is rich in polyunsaturated fatty acids (PUFAs), which have potential as therapeutic compounds for the treatment of neurodegenerative and cardiovascular disease. However, the effect of hempseed meal (HSM) intake on the animal models of these diseases has yet to be elucidated. In this study, we assessed the effects of the intake of HSM and PUFAs on oxidative stress, cytotoxicity and neurological phenotypes, and cholesterol uptake, using Drosophila models. HSM intake was shown to reduce H₂O₂ toxicity markedly, indicating that HSM exerts a profound antioxidant effect. Meanwhile, intake of HSM, as well as linoleic or linolenic acids (major PUFA components of HSM) was shown to ameliorate Aβ42-induced eye degeneration, thus suggesting that these compounds exert a protective effect against Aβ42 cytotoxicity. On the contrary, locomotion and longevity in the Parkinson’s disease model and eye degeneration in the Huntington’s disease model were unaffected by HSM feeding. Additionally, intake of HSM or linoleic acid was shown to reduce cholesterol uptake significantly. Moreover, linoleic acid intake has been shown to delay pupariation, and cholesterol feeding rescued the linoleic acid-induced larval growth delay, thereby indicating that linoleic acid acts antagonistically with cholesterol during larval growth. In conclusion, our results indicate that HSM and linoleic acid exert inhibitory effects on both Aβ42 cytotoxicity and cholesterol uptake, and are potential candidates for the treatment of Alzheimer’s disease and cardiovascular disease.     

大豆耐旱种质鉴定和相关根系性状的遗传与QTL定位

从301份黄淮海和长江中下游地区代表性大豆地方品种和育成品种(系)中按根系类型选取59份,在苗期干旱胁迫和非胁迫条件下对地上部和地下部性状进行2年重复鉴定,发现材料间性状隶属函数值具有丰富遗传变异,以株高、叶龄、根干重和茎叶干重隶属函数的算术平均数为抗旱综合指标从中筛选出汉中八月黄、晋豆14,科丰1号,圆黑豆等强耐旱型(1级)和临河大粉青、宁海晚黄豆等干旱敏感型(5级)材料。比根干重、比总根长、比根体积与耐旱隶属函数平均值均呈极显著正相关,可作为耐旱性的根系性状指标。利用“科丰1号×南农1138 2”(1级×4级)衍生的RIL群体为材料,对耐旱相关根系性状采用主基因 多基因混合遗传模型分离分析法进行遗传分析并进行QTL定位。结果表明,该两亲本间比根干重、比总根长、比根体积的遗传均为两对主基因加多基因模型,后两者主基因间有连锁(重组率分别为4.30%和1.93%);主基因遗传率为62.26%～91.81%,多基因遗传率为2.99%～24.75%;耐旱相关根系性状各主要由1对主基因控制,另1对效应较小。QTL分析检测到5、3、5个QTLs分别控制比根重、比根总长、比根体积,位于N6 C2、N8 D1b W、N11 E、N18 K连锁群上。3性状各有1个贡献率大的QTL(Dw1,Rl1,Rv1),而且均位在N6 C2的STAS8_3T STAS8_6T相同距离的区段上,其他QTLs效应均较小。分离分析与QTL定位的结果相对一致。     

盐磷耦合胁迫下大豆的生长和钠、磷离子长距离运输

以2个耐盐性和磷效率有差异的大豆品种为材料,采用水培方法,探讨盐分与缺磷耦合胁迫对大豆生长和钠、磷离子长距离运输影响的结果表明:(1)盐分和低磷胁迫对大豆生长有交互作用,磷浓度相对较高(2MMOL·L-1)时大豆耐盐性降低;(2)钠由木质部的向顶部运输增加,钠在韧皮部的再分配增多;(3)盐胁迫下磷在木质部的运输能力提高,韧皮部中磷的再分配受影响不大;(4)磷盐互作对大豆生长的影响在品种之间无差异。     

Manganese and iron interactions on their uptake and distribution in soybean (Glycine max (L.) Merr.)

Summary The uptake and distribution of iron and manganese were studied in a manganese-sensitive soybean cultivar (‘Bragg’) grown over a range of supply levels of these nutrients in solution culture. At high (90 and 275 μM) manganese levels, increasing the iron concentration in solution from 2 to 100 μM partially overcame the effects of manganese toxicity. Interactions between manganese and iron occurred for dry matter yields, rate of Mn absorption by the roots, and the proportions of manganese and iron transported to the tops. No interaction was observed for the rate of root absorption of iron. The percentage distribution of manganese in the plant top increased with increasing iron, despite a reduced rate of Mn uptake. On the other hand, iron uptake was independent of solution Mn concentration and increased with increasing solution Fe. Also more iron was retained in the roots at high Mn and/or Fe levels in solution. Concentrations of manganese and iron in roots, stems and individual leaves were affected independently by the manganese and iron supplyi.e. without any interaction occurring between the two elements. In general, the concentration in a plant part was related directly to the solution concentration. Symptoms resembling iron deficiency correlated poorly with leaf Fe concentrations whereas high levels of manganese were found in leaves displaying Mn toxicity symptoms.     

Trifluralin-induced chlorosis in soybeans (Glycine max (L.) Merr.) grown on clayey,high pH soils

Summary The cause of leaf chlorosis, frequently observed on soybeans (Glycine max (L.) Merr.) grown on high pH soils of the Mississippi Blackland Prairie, is thought to be low Fe availability and restricted rooting. Three greenhouse experiments were conducted using two soils, Sumter, a Rendollic Eutrocrept and Okolona, a Typic Chromudert; nine soybean cultivars differing in Feefficiency; and trifluralin (α-α-α-trifluoro-2,6-dinitro-N, N-di-propyl-p-toludine). Trifluralin at rates greater than 0.56 kg/ha caused chlorosis which was more severe on the Sumter, a soil low in available Fe. Fe-efficient cultivars were more resistant to the chlorosis induced by trifluralin than the Fe-inefficient cultivars. It was concluded that the chlorosis is an Fe deficiency caused by reduced uptake. The herbicide-induced chlorosis can be avoided by proper dosage and placement of the herbicide.     

大豆查尔酮合成酶(CHS)基因的克隆、表达及其在雪莲提取液中的代谢产物分析

以中国大豆为材料,利用PCR方法克隆查尔酮合成酶（Chalcone synthase,CHS）全基因,采用SOE法克隆得到去掉内含子的查尔酮合成酶基因,核酸序列分析表明,该基因编码区长1170bp,编码390个氨基酸,与已报道的CHS的cDNA序列同源率达到97%。构建pET-GMCHS工程表达质粒,通过大肠杆菌E.coli BL21（DE3）高效表达系统表达大豆CHS。 通过12% SDS-聚丙烯酰胺凝胶电泳分析表明,获得了分子量在42.9KD的一条蛋白质特异表达带。液相色谱分析大肠杆菌E.coli BL21（DE3）高效表达系统在雪莲提取液中的代谢产物,样品和空白对照样对比,样品在273nm,3.0min出现新的吸收峰,质谱分析结果表明CHS利用雪莲提取液中代谢中间产物合成了新的黄酮类物质。