小麦矮缩病毒引起的植株矮化与赤霉素代谢的相关性分析

【目的】由异沙叶蝉(Psammotettix alienus)传播的小麦矮缩病毒病是近年来中国西北部麦区严重发生的小麦病毒病害之一。受侵染的小麦植株严重矮化,有效分蘖减少,产量损失严重。论文旨在明确小麦矮缩病毒(Wheat dwarf virus,WDV)侵染小麦植株后矮化症状形成与赤霉素代谢调控的关系,为该病害的防治打下基础。【方法】以小麦品种扬麦12为试验材料,以异沙叶蝉为传毒介体饲毒后转移到1叶期的健康幼苗(3头/株)上进行传毒,同时以无毒异沙叶蝉取食健康幼苗为对照。根据试验需要,不同时间取样备用。为保证试验的准确性,经PCR检测为阳性的作为处理组试验材料;采用间接酶联免疫吸附(ELISA)法,利用植物赤霉素(GA3)试剂盒测定分析侵染第21天取样的小麦叶片赤霉素含量;将带毒条沙叶蝉接种的小麦苗分为两个平行处理组,接种后第7天分别用GA3(浓度为50 mg·L-1)和H2O进行叶面喷施处理,每隔一周处理一次。以无毒叶蝉接种后长势一致的小麦苗作为对照组,根据株高统计结果分析外施赤霉素对受侵染小麦植株的表型变化;以山羊草(Aegilops tauschii)的内根-贝壳杉烯合成酶(ent-kaurene synthase-like 3,KSL3)的基因编码区序列为参考基因设计引物(KSL3-F:5′-ATGATGGTGAATCCGCCGC-3′;KSL3-R:5′-TTAATGGTTGATCTTTGTTT-3′),对扬麦12的KSL3进行克隆和序列分析;分别取接种后7、14和21 d的小麦植株叶片,提取RNA后反转录,以克隆得到的Ta KSL3基因序列设计引物(Ta KSL3-F:5′-GAGACATGTGCCATGGCGTTC-3′;Ta KSL3-R:5′-CGTGTCACTCAGATCGGTGGAG-3′),选择小麦翻译延伸因子1A(EF-1α)作为内参基因,利用荧光定量PCR方法分析赤霉素代谢相关基因的转录水平。【结果】经ELISA检测发现,接种21 d的发病植株赤霉素含量与健康植株相比降低了28.9%;通过施用浓度为50 mg·L-1的赤霉素后,发病植株的平均株高相比对照组显著增加35.9%;采用同源克隆得到了完整的小麦赤霉素合成途径关键酶KSL3的编码区序列,长度为1 827 bp,编码608个氨基酸,BLAST比对分析发现该DNA序列与山羊草KSL3编码区序列相似度为85.2%。经荧光定量检测发现受小麦矮缩病毒侵染后小麦KSL3表达量显著下降,接种14 d降低为对照组的35.7%,21 d降低为对照组的9.6%。【结论】小麦矮缩病毒的侵染导致赤霉素合成途径关键酶的表达量降低,可能使赤霉素合成受阻,赤霉素含量降低引发受赤霉素调节的细胞生物学过程异常,从而诱导矮化症状形成。研究结果为揭示小麦矮缩病毒侵染的致病机理和病害防控打下了基础。     

刺萼龙葵种子中适宜内参基因的筛选

入侵杂草刺萼龙葵Solanum rostratum Dunal传播扩散的主要载体是种子,研究其种子休眠萌发基因的激素调控对于其防除具有重要意义,而选择合适的内参基因可以提高相关基因表达分析的准确性。本研究以赤霉素、脱落酸和水处理的刺萼龙葵种子为材料,利用geNorm、NormFinder、BestKeeper和RefFinder 4种软件对15个候选内参基因进行表达稳定性评价,并通过检测ABI5(abscisic acid-insensitive 5)的表达验证所筛选的内参基因的适用性。结果表明,对于赤霉素、脱落酸和水处理过的种子,最稳定的内参基因分别为eIF(eukaryotic initiation factor)、SAND(SAND protein family)和ACT(β-actin);对所有种子样本而言,PP2Acs(a catalytic subunit of protein phosphatase 2A)是最稳定的内参基因。研究结果将为刺萼龙葵种子休眠萌发的遗传调控研究提供重要参考。     

Effects of forage type and gibberellic acid on nitrate leaching losses

Nitrogen (N) leaching from soil into water is a significant concern for intensively grazed forage‐based systems because it can cause a decline in water quality and is a risk to human health. Urine patches from grazing animals are the main source of this N. The objective of this study was to quantify the effect that forage type and gibberellic acid (GA) application had on N leaching and herbage N uptake from urine patches on perennial ryegrass–white clover (RGWC), Italian ryegrass and lucerne. A lysimeter study was conducted over 17 months to measure herbage growth, N uptake and N loss to water beneath each of the three forage types with the following treatments: control, urine (700 kg N/ha) and urine with GA (8 g GA active ingredient/ha). Compared with RGWC (205 kg N/ha), N leaching losses were 35.3% lower from Italian ryegrass (133 kg N/ha) and 98.5% higher from lucerne (407 kg N/ha). These differences in leaching loss are likely to be due to winter plant growth and N uptake. During the winter months, Italian ryegrass had higher N uptake, whereas lucerne had lower N uptake, compared with RGWC. The application of GA had no effect on N leaching losses, DM yield or N uptake of forage treated with 700 kg N/ha urine.     

赤霉素处理对大白菜阶段发育的影响

用赤霉素处理未通过春化阶段的大白菜，植株可以抽薹开花；赤霉素处理已通过春化阶段的大白菜，可以促进植株提前抽薹开花。大白菜品种不同，赤霉素处理的效果有差异。     

Photoperiodic Regulationof Apical Growth Cessation in Northern Tree Species

Summary

Tree species adapted to the climatic conditions of the northern boreal and subarctic vegetation zones have a capacity to develop a very high level of frost hardiness, even to survive the temperature of liquid nitrogen in midwinter. Proper timing of hardening, as well as of dehardening, is crucial for winter survival of these species. In northern tree species, cessation of apical elongation growth and bud set is a prerequisite for developmental and metabolic processes leading to hardening, and this chain of events is induced by photoperiod. The northern tree species are closely adapted to the local light climate and display photoperiodic ecotypes. The critical photoperiod is under genetic control and increases with increasing latitude of origin of the eco-type. The photoperiod is probably perceived by the phytochrome system, but the role of other pigment systems, like cryptochrome, has not been studied in woody plants. Phytochrome genes have been cloned from both conifers and deciduous species, but so far we do not have any information about possible differences between photoperiodic ecotypes at the phytochrome level. Northern and southern ecotypes have different responses to red:far red ratios, which could indicate differences in composition of their phytochrome systems, for example, the proportions of phytochrome A and B. Both phytochrome A and phytochrome B can be involved in photoperiodic responses. Experiments with transgenic hybrid aspen suggest that responses to photoperiod could be affected by the amount of phytochrome A present in plants. In deciduous species, the plant hormone gibberellin A₁ (GA1) can completely substitute for a long photoperiod, and short day induced cessation of growth is preceded by a significant reduction of GA1 levels, particularly in the elongation zone. Photoperiodic control of GA metabolism is supported by several studies, but very little is known about the interaction between phytochrome and GA metabolism and/or responsiveness to GA1. Although our knowledge is still very fragmentary, available results suggest that cessation of growth and initiation of hardening in trees can be controlled both through the phytochrome and the GA mediated systems. Research with tree species is a tedious and slow process, but with the emerging new methods and approaches, we may expect exciting new results in the near future.     

赤霉素对香荚兰豆荚生长的作用

西双版纳景洪地区网棚种植的香荚兰,年月连续喷施次不同浓度的赤霉素,间隔喷施一次,19997315d对豆荚都有不同程度的促长作用。其中喷施×6010-6的效果最佳,比对照(清水)豆长增长,豆粗增加6.7% ,一级豆率增加。3.1%63.4%     

植物生长调节剂在花期调控中的应用

利用人为的因素可控制花卉在自然花期之外开放,统称为花期调控。笔者主要综述了植物成花机理以及赤霉素等植物生长调节剂在花期调控中的机理和应用,以期对花期调控的应用起到借鉴作用。     

GA3介导miR171s及其靶基因VvSCLs调控葡萄种子发育的作用分析

【目的】鉴定vvi-miR171s成员及其靶基因,明确vvi-miR171s成员及其靶基因在葡萄种子发育中的重要作用及其响应赤霉素（Gibberellin,GA）调控种子发育的表达模式。【方法】以‘魏可’葡萄（‘Wink’）果实为试材,利用miR-RACE技术鉴定vvi-miR171a/b/e/f/g/h的成熟体序列;通过PsRNATarget软件预测vvi-miR171s的靶基因,利用生物信息学软件对其进行染色体定位、系统进化树、基因结构、保守结构域及motif分析;通过启动子作用元件分析预测vvi-miR171s及其靶基因的潜在功能;采用RLM-RACE和PPM-RACE验证vvi-miR171s对靶基因的裂解作用;利用qRT-PCR鉴定其应答外源GA₃在葡萄种子区中的表达模式。【结果】从‘魏可’葡萄果实中鉴定并克隆了6条葡萄vvi-miR171s成熟体序列,并用RLM-RACE和PPM-RACE鉴定到3条靶基因VvSCL6/15/22。靶基因的生物信息学分析显示,VvSCL6/22与苹果、桃子和樱桃同源基因遗传距离较近,而VvSCL15则与拟南芥、苹果、桃子和樱桃中的同源基因遗传距离相同,且VvSCLs的基因结构也与进化树中亲缘关系较近的基因相似;SCLs蛋白均具有GRAS结构域,鉴定到5个与GRAS结构域对应的保守motif,且它们含有的motif元件类型及排列顺序均相似,表明SCL家族结构较为保守。vvi-miR171s及靶基因的启动子均具有大量的GA、水杨酸（Salicylic acid,SA）和种子发育相关作用元件,表明它们可能参与响应GA、SA和种子发育过程;qRT-PCR表达分析显示,GA₃强烈抑制了种子区vvi-miR171a的表达,但同时显著上调种子区的VvSCL15和VvSCL22表达;vvi-miR171a和VvSCL15在对照组和GA₃处理后的种子/种子区均呈强烈的负相关,表明GA₃可能在葡萄种子/种子区显著增强vvi-miR171a对VvSCL15的负调控作用,从而介导葡萄种子发育。【结论】在‘魏可’葡萄中鉴定到vvi-miR171a/b/e/f/g/h 6个成员;均可裂解VvSCl6/15/22 3个靶基因;vvi-miR171a-VvSCL15可能作为主要的调控途径响应赤霉素并参与调控葡萄种子发育。     

不同激素处理对假酸浆种子萌发的影响

【目的】假酸浆因种子外被胶质严重影响种子的萌发,探究不同处理对假酸浆种子萌发的影响,增加其种子的萌发率。【方法】以假酸浆种子为试材,采用随机区组法,设置种子放置在纱布上 4 ℃低温处理,以及赤霉素（GA3）、6- 苄氨基腺嘌呤（6-BA）和萘乙酸（NAA）的 MS 培养基处理,定期观察并作记录。【结果】与常温对照相比,种子放置在纱布上 4 ℃低温处理显著促进假酸浆种子萌发,以处理 48 h 发芽率最高为 25 个百分点。MS 培养基添加激素处理中,10 mg/L GA3 处理假酸浆种子发芽率和发芽指数分别为 92.00% 和9.58,比 CK 分别提高 4.27 个百分点和 1.33;1 mg/L 6-BA 处理假酸浆种子发芽率最高、为 96.63%,2 mg/L 6-BA处理发芽势最高、为 73.33%,比 CK 分别增加 8.9 个百分点和 10.07 个百分点;1、2 mg/L NAA 处理假酸浆种子发芽率均为 88.86%,但发芽势和发芽指数均比 CK 低;与 CK 相比,10、50 mg/L GA3 处理假酸浆种子下胚轴长度显著增加,而 6-BA 和 NAA 处理的下胚轴长度显著降低。【结论】低温处理能够显著促进假酸浆种子萌发,MS 培养基添加低浓度激素处理均能提高假酸浆种子的发芽率,通过发芽率、发芽势、发芽指数和下胚轴长度综合比较,假酸浆种子的最佳激素处理为 MS+10 mg/L GA3。