SSR分子标记在作物遗传育种中的应用

SSR(simple sequence repeat)是建立在PCR技术上的一种广泛应用的分子标记,具有含量丰富、多态性高、共显性等优点。本文简要介绍了SSR分子标记技术的原理和特点,重点介绍了SSR分子标记技术在作物遗传育种中的应用,主要在作物遗传多样性、基因定位、分子辅助标记、遗传图谱构建、品种鉴定和纯度鉴定等方面进行阐述。     

5种杂交粳稻亲本SCAR标记的建立及其在真伪纯度鉴定中的应用

选用36个随机引物对"寒丰A"、"寒丰B"、"8204A"、"8204B"、"R161"等5份杂交粳稻亲本材料进行RAPD扩增,对其中特异RAPD标记片段进行克隆和测序.根据获得的特异DNA序列设计序列特征扩增区(SCAR)特异的引物,将18个RAPD标记转化成6个稳定的SCAR标记.用这些SCAR标记对亲本和杂种F1代单株进行检测,实验室检测种子纯度的结果与海南田间种植的结果基本一致.此外,应用水稻细胞质雄性不育特异的1对PCR引物,分辨出2对不育系/保持系亲本:"寒丰A"与"寒丰B"、"8204A"与"8204B".     

分子标记在作物育种中的应用

近20年来发展了多种分子标记,使作物育种学家有可能直接根据基因型而不只是表现型进行选择,在作物育种的各个方面具有重要的利用价值。本文简要综述了分子标记在作物育种方面的有关应用。主要内容有：（1）分子图谱构建与基因定位;（2）DNA指纹库的建立;（3）标记辅助选择;（4）F1杂种优势分析;（5）基于图谱克隆基因。     

利用标记基因检测转Bt基因棉种子纯度的研究

安徽农业大学植保系李长福、吴振廷、王学林等研究人员根据转Bt基因棉国抗 1号转入的外源基因中整合有GUS基因和NPTⅡ基因 ,建立了 3种种子纯度检测方法 ,即GUS组织化学染色法、NPTⅡ培养法和NPTⅡ叶片涂抹法 ,准确率达到 98% ,在检测转Bt基因棉原始群系 2 0 - 1时 ,与生物测定比较准确率同样在 98%。为此 ,这 3种方法可用转Bt基因棉扩大规模制种过程中和推广应用良种纯度的检测 ,相应方法同样适用于基因组中整合有GUS基因和NPTⅡ基因的其他转Bt基因棉的品种。不仅如此 ,还适用于携带同一Bt杀虫蛋白基因的所有转Bt基因植物。同时…     

DNA分子标记技术在玉米种子纯度鉴定中的应用

从常规鉴定、生化鉴定及分子标记技术鉴定等方面,阐述了玉米种子纯度鉴定的重要性。进一步对RAPD、RFLP、SSR和AFLP等分子标记技术在种子纯度鉴定和品种真实性分析中的应用潜力及存在问题进行比较,得出SSR分子标记技术是目前种子纯度和品种真实性鉴定中最适宜的技术。     

SRAP标记技术在甜瓜品种亲缘关系和种子纯度鉴定上的应用

利用SRAP分子标记对12份甜瓜材料进行亲缘关系鉴定和F1代杂交种进行纯度检测。结果表明:从40对引物中筛选出13对多态性好的引物,共扩增出78条带,多态性带的比例为50%;聚类分析结果显示,12份甜瓜材料的相似系数在0.263～0.921之间,可将该12份甜瓜材料分为2大类,进一步细分为5类。利用引物Me15-Em7和Me3-Em16对‘红绿早脆’100S杂交种进行纯度检测,杂交种纯度为77%。研究为甜瓜品种间亲缘关系及种子纯度检测提供了可靠的理论依据。     

利用微卫星标记鉴定杂交水稻冈优22种子纯度的研究

水稻是重要的粮食作物之一,早在70年代我国就已实行了杂交稻的三系配套,目前杂交水稻在我国已大面积种植。由于杂交稻种子的真伪无法从种子形态上鉴别,这给造假者以可乘之机。在正常的情况下,种子的纯度也会因质量控制不严而给农业生产造成大量损失。这些都迫切需要有一种…     

RAPD分子标记及其在作物遗传育种中的应用

ＲＡＰＤ分子标记是一种新型的分了遗传标记,其原理是采用１０个碱基的随机引物,以基因组ＤＮＡ为模板进行ＰＣＲ随机扩增。其优越性在于其方法简单;分析中的花费少,用时短;分析所需的ＤＮＡ用量也不多等。本文着重介绍以电泳技术和ＰＣＲ扩增技术为核心的分子标记以及在农作物遗传育种中的应用。     

陕油8号种子纯度的RAPD鉴定研究

从杂交油菜“陕油8号”及其亲本中提取基因组DNA,用100个RAPD随机引物进行扩增,从中筛选出3个可将亲本和子代区分的引物BA208、BA1090、BA497。BA208产生亲本互补的特征带BA208-1050bp、BA2081250bp;BA1090产生母本特征带BA1090-700bp,BA497产生父本特征带BA497-870bp,上述谱带均在子代中出现。以BA208产生的特征谱带作为分子标记对杂交油菜种子纯度鉴定得到了一致的结果,并与大田纯度检测结果一致。BA497可将“陕油8号”与当地4个主栽品种有效区分。此外,还对双引物共同鉴定杂交种子纯度问题进行了初步探讨。     

分子标记的种类及其在作物遗传育种中的应用

分子标记技术近年来发展很快,目前,常用的分子标记主要可以分为基于杂交的分子标记、基于PCR的分子标记、基于限制性酶切和PCR结合的分子标记以及新一代分子标记.本文对这几类分子标记中常用类型的基本原理进行介绍,并从分子标记在作物种质遗传多样性、基因定位和基因克隆以及分子标记辅助选择育种和分子设计育种中应用进行了阐述.     

Rodent seed predation and seedling recruitment in mesic grassland

Seedling recruitment of two grasses (Arrhenatherum elatius and Festuca rubra) and two herbs (Centaurea nigra and Rumex acetosa) was measured in areas with and without rodents to which seeds of each species were sown at three seed densities (1000, 10,000 and 50,000 seeds m⁻²) in two seasons (spring and autumn 1995). Seed removal was measured for 10-day periods and the fate of seedlings was followed for 15 months after sowing. The proportion of seed removed ranged from 6 to 85% and increased with increasing seed density for each species. Rodents had no effect on seedling emergence or survival in the spring sowing. In the autumn sowing, rodents reduced seedling emergence of all four species sown at 1000 and 10,000 seeds m⁻² but had no impact at 50,000 seeds m⁻², presumably because of microsite limitation. We suggest the difference between spring and autumn arose because emergence was seed limited in autumn but microsite limited in spring; microsite availability was higher in autumn because a summer drought killed plants, reduced plant biomass and opened up the sward. Fifteen months after the autumn sowing, fewer A. elatius and C. nigra seedlings survived on plots exposed to rodents. This result reflected not only the reduced seedling emergence but also increased seedling mortality (seedling herbivory) in sites exposed to rodents. In contrast, F. rubra and R.acteosa showed density-dependent seedling survival which compensated for initial differences in seedling emergence, so that no effect of rodents remained after 15 months. The results suggest that rodent seed predation and seedling herbivory exert strong effects on seedling recruitment of A.elatius and C. nigra when recruitment conditions are favourable (conditions that lead to high microsite availability) and may contribute to both species being maintained at low densities in the grassland. The results also demonstrate that highly significant impacts of rodent seed predation at the seedling emergence stage can disappear by the time of plant maturation. Received: 2 March 1998 / Accepted: 28 September 1998     

城市污泥对作物种子发芽及幼苗生长影响的初步研究

不同类型污泥及其堆肥对通菜、菜心种子发芽和幼苗生长影响的研究表明 ,深圳生污泥、佛山生污泥和广州生污泥及其堆肥对通菜种子的发芽率有明显的影响 ,前者仅为73% ,其余的在 85%左右 ,均低于对照处理 ( 93% ) .而它们对菜心种子的发芽率 ,除了深圳生污泥有抑制作用外 ( 75% ) ,其余均表现为促进作用 ( 90 % ) ,均高于对照处理 ( 85% ) .它们对于两种作物幼苗的生长则均表现为不同程度的抑制作用甚至是损害作用 .污泥及其堆肥中抑制作物种子发芽和幼苗生长的物质主要是有机酸和醛类等有机物 .生污泥经过消化作用或适当的堆沤作用可降解这些有机物 ,从而减弱或消除这种抑制作用 .因此 ,生污泥经过消化作用或适当的堆沤作用后用于农业生产更为安全 .但对刚施用污泥或污泥堆肥的耕地最好不要直接播种 .     

沙埋对六种沙生植物种子萌发和幼苗出土的影响

研究了沙埋对科尔沁沙地6种优势植物的种子萌发和幼苗出土的影响．进行0．2、4、6、8、10和12cm等7个深度沙埋处理．结果表明,在不同沙埋处理时,沙蓬萌发差异显著。而差不嘎蒿2锄埋深与其他埋深的发芽差异显著,其他4种植物0cm埋深与其他埋深的发芽差异显著;沙埋对所有植物幼苗出土均有显著影响,埋深增加,出苗率减小;繁殖体大的物种与繁殖体小的物种相比,能从更深沙层中出苗,幼苗出土最大深度排序为苦参>东北木蓼≥沙蓬>山竹子>雾冰藜>差不嘎蒿．     

Species identification of polygonati rhizoma in China by both morphological and molecular marker methods

Morphological markers as well as two types of molecular markers, inter-sample sequence repeat (ISSR) and start codon targeted (SCoT) are suitable for species identification of the polygonati rhizoma germplasms. In this paper, we adopted these methods for the identification of rhizomes collected from 47 areas in China. Based on their morphological characters, the collected germplasms were classified into two populations, one with alternate leaf arrangement and the other with verticillate leaf arrangement, and they were comprised of five species and fourteen subgroups. Of the five species identified: Polygonatum kingianum, P. cirrhifolium, P. alternicirrhosum, and P. sibiricum belonged to one cluster, and P. cyrtonema belonged to a different cluster. According to the analysis of both ISSR and SCoT markers, all germplasms with greater genetic similarity were classified into one group. Especially, P. sibiricum and P. cirrhifolium, which shared ～80% similarity, were clustered together, whereas the germplasms identified as P. kingianum with ～86% similarity formed a separate clade. P. kingianum showed a much greater genetic similarity with P. cyrtonema than with P. sibiricum. The multidimensional scaling analysis further verified the accuracy and reliability of the molecular marker-based results. Thus, both morphological and molecular methods should be combined for the differentiation of germplasms such as those of polygonati rhizoma.     

种子萌发和幼苗生长对沙丘环境的适应机制

综述了植物沙生适应机制的研究及其进展.一些植物的种子在刚成熟时具有休眠特性.种子需要适度沙埋促进萌发并实现幼苗定居,但过度沙埋则会抑制种子萌发和出苗.在沙层深处,没有萌发的种子会进入休眠状态,形成土壤种子库.幼苗通过增加节数和延长节间来适应沙埋.沙埋深度超过植物的忍耐限度会抑制幼苗生长,甚至导致幼苗死亡.沙生植物必须适应其他环境因子,如盐风与土壤盐分、昆虫采食、土壤养分亏缺等,才能在沙丘上成功生长.沙蚀可导致幼苗根系暴露并干燥脱水.一些沙漠植物的幼苗在萌发后可忍耐一段时期的干燥,水分条件得到满足之后,幼苗能够恢复生长.     

温室白粉虱SCAR分子鉴定技术的建立及应用

温室白粉虱Trialeurodes vaporariorum是为害我国蔬菜作物的重大害虫之一。本研究采用随机扩增多态性DNA(Random Amplified Polymorphic DNA,RAPD)技术对温室白粉虱进行研究,筛选出一条620bp的特异性片段(GenBank登录号为JQ690764),据此片段的测序结果设计一对特异性的序列特异性扩增区(Sequence characterized Amplified Region,SCAR)标记(Tv-F和Tv-R),可成功从室内饲养的和从不同地区田间采集的温室白粉虱的基因组DNA中扩增出一条412bp的特异性条带,而不能从供试的其他粉虱类害虫中扩增出该相应条带。单头温室白粉虱成虫的基因组DNA稀释1000倍时,该SCAR标记仍可成功扩增出预期条带,显示出极高的灵敏度。该SCAR标记对温室白粉虱的基因组DNA扩增重复性和稳定性好,且操作简便,灵敏度高,可用于样品的大规模检测,为田间温室白粉虱的快速识别鉴定及其有效防治提供了技术基础。     

A new mechanism in plant engineering: The potential roles of microRNAs in molecular breeding for crop improvement

MicroRNAs (miRNAs) are small, endogenous, noncoding RNAs that negatively modulate the expression of genes by inhibiting translation or by promoting the degradation of target mRNAs. miRNAs are now known to have greatly expanded roles in a variety of plant developmental processes, in signal transduction, and in the response to environmental stress and pathogen invasion. Because of their ability to inactivate either specific genes or entire gene families, artificial miRNAs function as dominant suppressors of gene activity when brought into a plant. Consequently, miRNA-based manipulations have emerged as promising new approaches for the improvement of crops. This includes the development of breeding strategies and the genetic modification of agronomic traits. Herein, we highlight new findings regarding the roles of miRNAs in plant traits, and describe the current miRNA-based plant engineering approaches. Finally, we consider the feasibility of modulating current approaches to address future challenges such as breeding programs to increase crop yield.     

胁迫时间与非毒性离子对重金属抑制拟南芥种子发芽及幼苗生长的影响

测定了Hg²⁺、Cd²⁺、Cu²⁺、Pb²⁺单一重金属胁迫对拟南芥种子发芽和幼苗生长的影响.结果表明,重金属对幼苗生长的毒性大于对种子发芽的毒性,以抑制种子发芽的IC₅₀为指标,4种重金属的毒性顺序为Hg²⁺＞Cd²⁺＞Pb²⁺/Cu²⁺,以幼苗生长为指标,则毒性顺序为:Cu²⁺＞Hg²⁺＞Cd²⁺/Pb²⁺,并随着胁迫时间延长,种子萌发率下降.此外,不同重金属在不同发芽时段对种子的毒性也不尽相同,Cd²⁺的毒性在种子吸水后的0～12 h大于12～24 h,而Hg²⁺毒性在12～24 h大于0～12 h,其中,种皮对减轻重金属毒性起着十分重要的作用.通过非毒性离子(Ca²⁺、Mg²⁺、K⁺、Na⁺)与重金属离子(Hg²⁺、Cd²⁺、Cu²⁺、Pb²⁺)交互作用对拟南芥种子发芽及幼苗生长效应的研究发现, mmol·L^-1的Ca²⁺、Mg²⁺、K⁺、Na⁺可以增强Hg²⁺对种子发芽的毒性,但对Cd²⁺的毒性却没有影响.对于幼苗来说,Ca²⁺、Mg²⁺、K⁺、Na⁺可以显著增强Hg²⁺的毒性,Ca²⁺可以缓解Cd²⁺的毒性,但却增加Cu²⁺的毒性,K⁺可以缓解Pb²⁺对幼苗的毒害作用.最后,本文对重金属的毒害机理进行了探讨.