基因芯片技术在植物研究中的应用

简述了基因芯片的原理、特点、分类及制作方法,同时详细综述了基因芯片技术在植物研究中的应用。     

DNA序列在植物系统学研究中的应用

植物DNA序列由于进化速率上的差异而适用于不同分类阶元的系统发育研究,因此,针对某一特定的系统学问题选择相应合适的分子片段是分子系统学研究中最为关键的一步。在前人研究的基础上,主要讨论了目前分子系统发育和进化研究中一些常用的DNA序列的适用范围,包括nrDNA的18S基因及ITS等非编码区,cpDNA的编码基因（rbcI、matK、ndhF、atpB）及非编码区序列（rpL16、rpoC1、rps16、trnL-F和trnT-L）和应用较少的mtDNA。研究表明,18S、rbcI等编码基因及mtDNA一般适用于较高分类阶元甚至整个种子植物谱系间的系统发育的探讨,而ITS及cpDNA的非编码区序列等因其较快的进化速率多用于较低分类阶元的系统关系研究。     

现代生物技术在植物研究中的应用

伴随着生物科学世纪的到来,生物技术广泛应用于农业、医药、轻工业、食品、环保、海洋和能源等方面。列举了几项现代生物技术在植物学研究领域中的应用,从几个角度揭示了现代生物技术在人们日常的生产生活中所扮演的重要角色。     

双链RNA技术在植物病毒研究中的应用

含ＲＮＡ基因组的植物病毒在复制时会产生双链ＲＮＡ（ｄｓＲＮＡ）。本文介绍了用ＣＦ－－１１纤维素粉提取ｄｓＲＮＡ的步骤,并列举了ｄｓＲＮＡ在植物病毒研究中的应用,其主要应用有（１）用于检测植物病毒;（２）用于病毒株系分化;（３）用于病毒卫星ＲＮＡ及亚基因组ＲＮＡ研究;（４）用于侵染性测定及病毒基因组功能研究等。     

基因枪技术在植物基因工程研究中的应用

文章介绍了近年来采用基因枪技术在植物基因工程研究中开展的一些工作。     

酵母双杂交技术及其在植物研究中的应用

酵母双杂交技术能有效地检测蛋白质之间的相互作用.该文简单介绍了此技术的工作原理和操作程序,以及十几年来此项技术在植物功能基因组学和蛋白组学研究中的应用情况.     

mRNA差别显示技术在植物发育研究中的应用

mRNA差别显示技术是近年发展起来的研究基因表达差异的有效方法,它不仅可比较基因表达差异、追踪已知基因的表达情况,而且可作为寻找特异表达基因的重要手段。本文介绍此方法在植物春化作用、光周期反应、花发育、雄性不育、成熟、衰老、种子发育和休眠及根发育等研究领域的应用概况。     

分子标记的发展及其在植物研究中的应用

概述了分子标记的研究进展 ,并对其在植物研究领域中的应用及前景作了介绍和展望 .     

基因组学方法在植物抗逆性研究中的应用

由于植物抗逆性遗传极其复杂,因而植物抗逆性能（包括抗非生物胁迫如盐碱,干旱,低温等的能力和抗生物协迫如真菌,细菌,病毒和线虫的能力等）的提高受到了极大限制,近年来,基因组学的兴起对我们全面理解植物抗逆性起着革命性作用,结果基因组学将会使我们挖大量全新的抗逆基因,并能揭示各抗逆性基因的详细结构以及抗逆性遗传进化机理,功能基因组学将会阐明植物抗逆中的复杂的调控,网络,揭示涉及抗逆蛋白的多样性,通过比较基因学的研究,可以把从模式植物上获得的抗逆遗传信息推广到基因组较复杂的植物上去,大规模的全新基因的发现及其在抗逆反应中的表达模式的研究和它们在抗逆应中作用的理解将会利用遗传工程进行植物抗逆育种提供广阔的前景。     

PCR技术在植物病原细菌研究中的应用

利用rep-PCR,AFLP和RAPD等多种基因组指纹分析方法,指纹图谱结合计算机辅助分析,用于研究植物病原细菌群体遗传多样性;遗传多样性图谱有助于理解病原细菌的分类,群体结构。还有利于设计特异,灵敏,快速检测策略用于植物病原检测和病害诊断。已有许多以PCR为基础的检测技术如rDNA-PCR,ITS-PCR,ARDRA等,有很多特异性引物及检测程序,已在植物病原检测和病害诊断中广泛应用,PCR技术的应用和计算机的辅助分析。为植物病原细菌群体动态和生态学知识提供了基本框架,使植物病理学进入一个新时代。     

Fine structure of aphid stylet pathways and its use in host plant resistance studies

Conclusion Although there have been reports, based on light microscope observations, of damage to mesophyll tissue as a result of stylet penetration, we saw no evidence of this in our wax sections. However, the use of the electron microscope revealed that such damage does occur, and has also shown that both inter- and intracellular penetration routes exist, often within the same track, whereas we had formerly believed the stylet pathways of these aphids to be almost entirely intercellular. The intramural-extracellular route of penetration, characterised by the presence of stylets and or saliva between the cell wall and plasmalemma, requires the greater resolution of electron microscopy and cannot be distinguished in light microscope preparations.Our results suggest that an accurate indication of the stylet pathway cannot be obtained from the use of light microscopy alone, and raise serious doubts about the value of previous studies describing the route of stylet penetration. Light microscopy shows the track only at the tissue level and not at the cellular level. We advocate that all future studies of aphid stylet penetration should utilise both light and electron microscope studies because only the higher resolution of the latter technique will indicate the true stylet pathway and the end points of the tracks. Only then can the feeding site be determined with any precision.     

Analysis of resistance in tomato and sweet pepper against the greenhouse whitefly using electrically monitored and visually observed probing and feeding behaviour

Different types of plant resistance against the greenhouse whitefly, Trialeurodes vaporariorum, were distinguished according to the tissue location of the resistance factors. The effects of resistance factors were compared by measuring the electrically monitored and visually observed probing and feeding behaviour of whiteflies on two resistant tomato genotypes (82207 and 82216), a susceptible tomato cultivar, and a non-host plant, sweet pepper. On sweet pepper, whiteflies displayed very short first probes, very long pathway probing and spent much time on non-feeding activities such as walking and standing still. Also, a high percentage of whiteflies rejected sweet pepper without ingesting substances from the phloem vessel. These data suggest a strong resistance that is based on the factors present in surface/epidermis and/or mesophyll layers of this plant. The behaviour of whiteflies on tomato 82207 was very different from that on sweet pepper. On tomato 82207 whiteflies apparently did not perceive resistance factors on the leaf surface and in the mesophyll. Resistance factors appeared to be present in the phloem tissue, because a higher number of phloem phases, longer phloem salivation periods and shorter phloem ingestion periods were observed when compared with the susceptible tomato cultivar. Tomato 82216 showed no clear resistance factor in the phloem. The importance of combined EPG and behavioural research for host-plant resistance studies are discussed.     

Insect feeding on cucumber mediated by rhizobacteria-induced plant resistance

Select strains of plant growth-promoting rhizobacteria (PGPR) were evaluated in greenhouse experiments with cucumber for induction of resistance against cucumber beetle (Diabrotica undecimpunctata howardi Barber) feeding and the beetle-transmitted cucurbit wilt disease. When beetles were given a choice between PGPR-treated and nontreated cucumber, their feeding on stems and cotyledons and the severity of wilt symptoms were significantly lower on PGPR-treated plants. HPLC analysis demonstrated that cotyledons from PGPR-treated plants contained significantly lower concentrations of the cucumber beetle feeding stimulant cucurbitacin than nontreated plants. These results suggest that a mechanism for PGPR-induced resistance against cucumber beetle feeding may involve a change in the metabolic pathway for cucurbitacin synthesis.     

Probing behavior of the tea green leafhopper on different tea plant cultivars

The probing behaviors of the tea green leafhopper, Empoasca vitis (Gothe), on 9 tea cultivars were studied using video-text Direct Current-Electrical Penetration Graph, i.e., DC-EPG. The following 7 types of waveforms produced by the leafhopper stylet probing were determined: A, stylet pathway formation; S, salivation when stylets pierce into and stay in phloems; C, active ingestion before stylets reach phloems; E and F, passive ingestion in phloems; R, the insect resting with its stylet inserted into the leaf tissue and NP without probing. The 9 tested tea cultivars were categorized into 3 groups by the cluster analysis according to the number of probes per insect, waveform durations, or duration per probe of various waveforms on different tea cultivars. Waveforms S, E and F correlated to the main feeding activity of the leafhopper and may provide valuable information on predicting the resis-tance level of the tea plants to the leafhopper. The resistance level of the 9 tea cultivars to the leafhopper was ranked based on the durations of waveforms S, E and F, as well as the duration per probe including various waveforms. The ranking order of the resis-tance was: Longjingchangye > Hangdan > Zhenghedabaicha > Qianmei 601 > Hongyafoshuo > Zhongcha 102 > Zhongcha 302 > Longjing 43 > Anjibaicha, which corresponded to the resistance level determined by the population density (infestation) of the leaf-hopper on the 9 tea cultivars in the tea fields. Our study suggests that this simple and convenient DC-EPG technique might have great potential as a reliable tool to predict the resistance of tea cultivars to the tea leafhopper.     

Probing behavior of the tea green leafhopper on different tea plant cultivars

The probing behaviors of the tea green leafhopper, Empoasca vitis (Gothe), on 9 tea cultivars were studied using video-text Direct Current-Electrical Penetration Graph, i.e., DC-EPG. The following 7 types of waveforms produced by the leafhopper stylet probing were determined: A, stylet pathway formation; S, salivation when stylets pierce into and stay in phloems; C, active ingestion before stylets reach phloems; E and F, passive ingestion in phloems; R, the insect resting with its stylet inserted into the leaf tissue and NP without probing. The 9 tested tea cultivars were categorized into 3 groups by the cluster analysis according to the number of probes per insect, waveform durations, or duration per probe of various waveforms on different tea cultivars. Waveforms S, E and F correlated to the main feeding activity of the leafhopper and may provide valuable information on predicting the resis-tance level of the tea plants to the leafhopper. The resistance level of the 9 tea cultivars to the leafhopper was ranked based on the durations of waveforms S, E and F, as well as the duration per probe including various waveforms. The ranking order of the resis-tance was: Longjingchangye > Hangdan > Zhenghedabaicha > Qianmei 601 > Hongyafoshuo > Zhongcha 102 > Zhongcha 302 > Longjing 43 > Anjibaicha, which corresponded to the resistance level determined by the population density (infestation) of the leaf-hopper on the 9 tea cultivars in the tea fields. Our study suggests that this simple and convenient DC-EPG technique might have great potential as a reliable tool to predict the resistance of tea cultivars to the tea leafhopper.     

The combination of electronic monitoring and video-assisted observations of plant penetration by aphids and behavioural effects of polygodial

Simultaneous electronic and close-up video recordings were made of behaviour during the initial 15 min of plant contact by adult, apterous Aphis fabae Scopoli on tick bean seedlings (Vicia faba Moench). Electronic techniques accurately determined stylet penetration of plant tissue and there was a close correlation between penetration and periods during which the insect antennae and body were immobile (r=0.994, n=60). Video techniques were then used alone to infer stylet penetration and the behaviour of aphids after various treatments was monitored. In particular, the time to first penetration, the number of penetrations, the mean duration of penetrations and the total time of penetration were observed. Behavioural differences were recorded between tethered (as required for electronic recording) and freelymoving insects as well as between fed and starved insects. The behaviour of starved aphids placed on beans treated with the plant-derived antifeedant, polygodial could not be distinguished from aphids on solvent-treated control beans. However, there were significant differences in behaviour of aphids which had previously been exposed to polygodial on plant or green/yellow paper surfaces for 24 h when compared with insects exposed to solvent alone. The possible modes of action of polygodial are discussed.     

Some physiological and biochemical aspects of plant resistance to cadmium effect. I. Antioxidative system

The hitherto studies on the effect of cadmium on plants have shown that it causes intensification of two types of unfavorable processes in plants: inactivation of macromolecules and cellular structures and induction of oxidative stress. In response, the plant organism activates processes restoring its homeostasis, i.e. those which remove reversible and irreversible changes. In removing the former a particular role is played by the antioxidative system containing enzymatic and nonenzymatic antioxidants, and removing the active forms of oxygen. The strategy of stress tolerance plays an important role in the plant resistance to cadmium and other toxic metals.     

青藏高原高寒草甸门源草原毛虫取食偏好及其与植物C、N含量的关系

草食性昆虫对不同植物物种的取食存在偏好,这种取食偏好可能受其自身对蛋白质和碳水化合物的需求及二者平衡的调节。以青藏高原高寒矮嵩草草甸31种常见植物及门源草原毛虫为对象,通过饲喂实验,研究了草食性昆虫对不同物种和不同功能群植物的取食偏好,及其与植物叶片C、N含量和C∶N之间的关系。在31种植物中,门源草原毛虫对19种植物进行了取食,尤其对矮嵩草、红棕薹草、藏异燕麦和垂穗披碱草四种植物表现出强烈的取食偏好,而对另外12种植物未进行任何取食。在物种水平上,门源草原毛虫取食量与植物叶片N含量呈显著负相关,与叶片C∶N呈正相关。从功能群水平上看,门源草原毛虫对莎草类的取食偏好最大,而对豆科植物取食偏好最低;相应地,莎草类植物叶片N含量最低、C∶N最高,而豆科植物叶片N含量最高、C∶N最低。因此,即使在土壤有效氮匮乏、植物生长受氮素限制的高寒草甸生态系统,植物体内N含量的增加也可能不利于草食性昆虫的取食。门源草原毛虫对优势植物矮嵩草和垂穗披碱草的取食对高寒矮嵩草草甸物种共存和生物多样性维持可能具有重要的作用。     

Effect of bioregulator-treated sorghum on greenbug fecundity and feeding behavior: implications for host-plant resistance

Three commercial and six experimental plant growth bioregulators were surveyed for their effect on aphid reproduction when applied to sorghum. Only CCC and PIX had a significant effect on the greenbug, Schizaphis graminum. Application of the commercial bioregulators CCC and PIX^R caused about a 50% decrease in aphid reproduction rate when applied to greenbug susceptible sorghum but had little effect when applied to a greenbug resistant sorghum line. Electronic monitoring of aphid probing behavior on CCC treated, greenbug-susceptible sorghum showed a response pattern which was indistinguishable from that normally observed on greenbug resistant lines and was different from that associated with aphid probing behavior on untreated susceptible lines. The isolated pectin content of the CCC treated susceptible sorghum was twice that of the controls and had twice the methoxy content. These results support the argument that pectin is a barrier to aphid-stylet penetration for phloem feeding aphids which probe intercellularly and that manipulation of pectin content and/or structure can be a major factor in host-plant resistance to sap-sucking insects.     

酵母双杂交在植物功能基因组研究中的应用

扼要介绍了酵母双杂交技术的原理,详细评述了该技术在植物功能基因组中的研究进展,并结合自己研究领域对该技术在植物领域发展方向作了展望。