桃蚜取食行为对大气CO_2浓度升高的响应

【目的】近年来随着人类活动的增加,温室气体尤其是大气CO_2浓度升高造成的虫害爆发已成为国际上关注的焦点,因此,研究拟南芥Arabidopsis thaliana上桃蚜取食行为的变化对大气CO_2浓度升高的响应意义重大。【方法】本研究以拟南芥和绿色桃蚜Myzus persicae为研究对象,利用野生型拟南芥Col-0,茉莉酸途径信号传导缺失突变体(jar1)、水杨酸途径信号传导缺失突变体(npr1)、乙烯途径信号传导缺失突变体(ein2-5)为材料,以大气CO_2浓度升高为影响因子,利用刺吸式电位仪(EPG)记录了桃蚜在不同处理的拟南芥上的取食波形。【结果】研究结果发现:CO_2浓度升高缩短了Col-0和jar1植株上蚜虫首次刺探时间和首次到达韧皮部的时间,却延长了npr1和ein2-5上蚜虫首次到达韧皮部的时间,降低了jar1植株上蚜虫总的刺探时间且增加了其总的取食韧皮部时间,但没有改变其它基因型植株上蚜虫总的刺探和取食时间;同时增加了野生型植株上蚜虫的刺探频率,却没有影响其它基因型植株上的刺探频率。【结论】CO_2浓度升高降低了野生型植株和jar1植株抗性,有利于蚜虫到达韧皮部;却增加了npr1和ein2-5上的植物抗性,从而不利于蚜虫到达韧皮部。     

多胺生物合成抑制剂D-精氨酸对拟南芥幼苗根系生长的影晌

为探讨多胺生物合成抑制剂D-精氨酸（D-arginine,D-Arg）对拟南芥根系生长的影响,首先用腐胺（0．1mmol‘L-1）和D—Arg（1．0mmol·L-1）处理种子萌发后生长2d的拟南芥幼苗。腐胺（0．1mmol·L-1）显著促进主根伸长,D-Arg（1．0mmol-L-1）显著抑制主根伸长,并对主根根尖的细胞形态有明显影响。为了进一步了解D—Arg影响拟南芥主根生长的机理,采用浓度梯度D．Arg处理幼苗根系。实验结果表明,随着D-Arg浓度增加（0．2～1．0mmol·L-1）,拟南芥幼苗主根生长受抑制的程度越严重。微分干涉观察主根根尖发现,外源施加D—Arg,引起拟南芥主根根尖分生区的细胞数目减少,使拟南芥幼苗表现出主根的伸长生长变缓。当分生区数目较少时,出现主根几乎不再仲长的现象。由此推测,多胺生物合成抑制剂D-Arg对拟南芥幼苗根生长的抑制作用机制,是D-Arg影响了其根尖分生区的细胞分裂活动,使分生区细胞数目减少,从而引起分生区长度减小,最终导致拟南芥主根仲长生长受到抑制。     

异三聚体G蛋白在NAA诱导的拟南芥根生长发育中的作用

以拟南芥的野生型(ws)、异三聚体G蛋白α亚基基因GPA1缺失突变体(gpa1-1,gpa1-2)和超表达突变体(wGα,cGα)为材料,通过施加不同浓度(0~0.2 mg/L)的NAA处理,对拟南芥根生长发育的一些形态指标进行了观测比较.结果表明:(1)随着培养基中NAA浓度的不断升高,5种基因型主根的伸长生长均受到抑制,且抑制作用随浓度升高而增强;4种突变体和野生型主根的生长在相同浓度NAA处理下,无明显差异;(2)NAA在一定浓度范围内,对拟南芥侧根的生长发育起促进作用;在NAA诱导的侧根生长中,G蛋白超表达突变体比野生型更敏感,缺失突变体则不敏感.初步证明G蛋白不参与主根生长发育的调节,而在侧根生长发育中可能起正调节作用.     

拟南芥根系发育的分子机制研究进展

拟南芥初生根和次生根的发育受不同遗传通路所调控,其中内源激素途径尤其是生长素途径在拟南芥主根、侧根以及根毛的发育过程中均发挥着重要作用.同时也存在一些不依赖于激素通路的遗传途径,如UPB1能通过调节根尖分生区和伸长区活性氧种类的平衡来调控根系顶端分生组织活性,进而影响根系的生长.本文对近年来国内外有关模式植物拟南芥根系发育的分子机制研究进展分别从初生根发育、侧根发育和根毛发育3个方面进行综述.     

尿黑酸对拟南芥酪氨酸降解缺陷突变体sscd1的影响

为了了解尿黑酸对拟南芥酪氨酸降解途径的影响,探讨植物中酪氨酸降解途径在长日照下是否被抑制,本研究以拟南芥野生型Col-0和酪氨酸降解途径缺陷突变体sscd1、hgo为实验材料,通过在培养基添加尿黑酸处理,分析长、短日照下尿黑酸对幼苗生长的影响,同时采用q RT-PCR分析尿黑酸对酪氨酸降解途径关键基因HGO、MAAI和SSCD1表达的影响。结果发现:在短日照下,尿黑酸处理能够加速酪氨酸降解途径缺陷突变体sscd1的死亡;在长日照下,尿黑酸也能导致sscd1突变体的死亡;尿黑酸处理上调拟南芥野生型、酪氨酸降解缺陷突变体sscd1中MAAI和HGO的表达,其中MAAI表达在sscd1突变体中的上调幅度更大。以上结果说明,外源尿黑酸处理确实可以激活植物体内酪氨酸降解途径,而且植物中酪氨酸降解途径在长日照下不会被抑制。此外,研究结果进一步证明酪氨酸降解途径中间代谢产物的积累可导致sscd1突变体的死亡。     

Rabdosinate调节生长素极性运输蛋白PIN1、PIN3和PIN4抑制拟南芥幼苗根生长

利用3种拟南芥生长素极性运输外运载体突变体及4种转基因株系研究了二萜rabdosinate抑制拟南芥幼苗主根及侧根生长的作用机制。结果显示,60~80 μmol·L^-1的rabdosinate显著抑制野生型拟南芥幼苗主根生长及侧根形成,而对突变体pin1、pin2和pin3主根未显示明显的抑制效应,对侧根的抑制减弱;发现rabdosinate （60~80 μmol·L^-1）引起生长素报告株系根尖DR5活性升高,并增加融合蛋白PIN1-GFP丰度以及减少PIN3-GFP和PIN4-GFP的丰度。推断rabdosinate可通过增加PIN1丰度促进了根部生长素向顶运输,而减少PIN3丰度降低根尖部生长素的横向转运,引起了生长素在根尖部的累积及生长素浓度梯度的改变,进而抑制幼苗主根生长及侧根发育。     

拟南芥铵超敏感突变体amosd和vtc1对外源铵的响应

分析了不同外源铵浓度(0、1、5、10、20mmol.L-1)处理下,2个铵超敏感突变体amosd和vtc1对于外源铵处理的响应差异。结果表明,尽管amosd和vtc1都表现为对外源铵超敏感,但二者对外源铵处理浓度的敏感性上存在差异。随着外源铵浓度的增加,vtc1比amosd先表现出铵中毒症状,更高浓度(20mmol·L-1)铵处理时amosd受到的毒害程度表现更加严重,AMOSD遗传位点的缺失容易导致植物出现铵毒害死亡。其次,二者在遭受外源铵胁迫时表现的最敏感部位有所不同,主要的毒害特征上存在差异,amosd在铵胁迫下首先表现在叶片尤其是新叶的发育受阻,而vtc1则主要表现在根部尤其是主根的伸长受阻。通过分区供应实验证明,amosd主要对于地上部供铵处理超敏感,对根部供铵处理不表现超敏感特性;而vtc1则相反,对根部供铵处理超敏感,对地上部供铵不表现超敏感特性。由此可见,amosd和vtc1这2个铵超敏感突变体在拟南芥铵毒害范围和部位上存在较大差异,与vtc1有所不同,amosd是一个叶源铵超敏感型突变体。在农业机械化叶面喷施施肥日益增加和环境铵沉降日益严重的当下,叶源型铵超敏感突变体amosd的获得为揭示植物地上部铵毒害机制提供了一个理想的遗传材料,对系统全面认识植物铵毒害机制,提高作物耐铵性状具有十分重要的意义。     

重力钝感的水稻突变体在重力和微重力条件下淀粉体的观察分析

植物向重性的研究一直受到关注,主要的研究集中在双子叶模式生物拟南芥中,而对单子叶植物的研究却很少。植物对重力感受的方式存在多种解释,但目前大量证据表明淀粉体—平衡石理论较为合理,它认为淀粉体作为平衡石在植物向重性反应中发挥了重要的作用。经过100多年的研究,现已从生理学与遗传学的角度证实了含有淀粉体的根冠中柱细胞和茎的内皮层细胞是植物重力感受的部位,淀粉体作为重力感受器被越来越多的实验证据证明。地球上重力无处不在,要研究微重力对植物体极性生长的影响只能借助于能模拟失重环境的回转器。近年来,人们对植物向重性机制的了解主要来自缺失或缺少     

PLDα1介导ABA调控的拟南芥主根伸长并参与根毛生长

探讨了磷脂酶Dα1（PLDα1）在ABA抑制拟南芥主根伸长过程中的作用。PLOα1基因突变体pldα1主根伸长受ABA抑制小于野生型（WT）;根系PLDα1活性在ABA处理下升高;拟南芥根细胞原生质体中活性氧（ROS）含量在ABA处理下升高,但是pldα1升高小于WT;根系NADPH氧化酶活性在ABA处理下升高,pldα1升高小于WT,外源加入10μmol/L^-1 PA（磷脂酸,PLD水解产物）后,前者活性显著升高;外源加入H2O2可诱导WT和pldα1主根伸长都受到抑制,且二者差异不明显。结果表明,PLDα1产生的PA通过激活NADPH氧化酶产生ROS介导ABA调控的拟南芥主根伸长过程。此外,初步探讨了PLDα1在拟南芥根毛尖端生长中的作用：pldα1突变体根毛长度小于WT,根毛尖端ROS和Ca^2＋浓度低于WT。     

植物根系向地性感应的分子机理与养分吸收

植物根系向地性是决定根系空间生长趋势的主要因素之一,对于养分吸收具有重要影响.认识根系向地性感应和根系生长变化的分子机理及其与养分吸收的关系,可为遗传改良根系性状、提高植物养分吸收效率提供理论依据.本文从重力感应、信号转导和生长素非对称分布等方面总结了植物根系向地性感应的分子机理,探讨了根系在养分胁迫下(特别是磷胁迫下)向地性变化的生理基础及其与养分吸收(特别是磷吸收)的关系,最后对根系向地性研究的若干问题进行了展望.     

Disruption of the F-actin cytoskeleton limits statolith movement in Arabidopsis hypocotyls

The F-actin cytoskeleton is hypothesized to play a role in signal transduction mechanisms of gravitropism by interacting with sedimenting amyloplasts as they traverse statocytes of gravistimulated plants. Previous studies have determined that pharmacological disruption of the F-actin cytoskeleton with latrunculin B (Lat-B) causes increased gravitropism in stem-like organs and roots, and results in a more rapid settling of amyloplasts in the columella cells of Arabidopsis roots. These results suggest that the actin cytoskeleton modulates amyloplast movement and also gravitropic signal transduction. To determine the effect of F-actin disruption on amyloplast sedimentation in stem-like organs, Arabidopsis hypocotyls were treated with Lat-B and a detailed analysis of amyloplast sedimentation kinetics was performed by determining amyloplast positions in endodermal cells at various time intervals following reorientation. Confocal microscopy was used to confirm that Lat-B effectively disrupts the actin cytoskeleton in these cells. The results indicate that amyloplasts in hypocotyl endodermal cells settle more quickly compared with amyloplasts in root columella cells. F-actin disruption with Lat-B severely reduces amyloplast mobility within Arabidopsis endodermal statocytes, and these results suggest that amyloplast sedimentation within the hypocotyl endodermal cell is F-actin-dependent. Thus, a model for gravitropism in stem-like organs is proposed in which F-actin modulates the gravity response by actively participating in statolith repositioning within the endodermal statocytes.     

Ammonium-induced loss of root gravitropism is related to auxin distribution and TRH1 function, and is uncoupled from the inhibition of root elongation in Arabidopsis

Root gravitropism is affected by many environmental stresses, including salinity, drought, and nutrient deficiency. One significant environmental stress, excess ammonium (NH(4)(+)), is well documented to inhibit root elongation and lateral root formation, yet little is known about its effects on the direction of root growth. We show here that inhibition of root elongation upon elevation of external NH(4)(+) is accompanied by a loss in root gravitropism (agravitropism) in Arabidopsis. Addition of potassium (K(+)) to the treatment medium partially rescued the inhibition of root elongation by high NH(4)(+) but did not improve gravitropic root curvature. Expression analysis of the auxin-responsive reporter gene DR5::GUS revealed that NH(4)(+) treatment delayed the development of gravity-induced auxin gradients across the root cap but extended their duration once initiated. Moreover, the β-glucuronidase (GUS) signal intensity in root tip cells was significantly reduced under high NH(4)(+) treatment over time. The potassium carrier mutant trh1 displayed different patterns of root gravitropism and DR5::GUS signal intensity in root apex cells compared with the wild type in response to NH(4)(+). Together, the results demonstrate that the effects of NH(4)(+) on root gravitropism are related to delayed lateral auxin redistribution and the TRH1 pathway, and are largely independent of inhibitory effects on root elongation.     

Conservation of the Plastid Sedimentation Zone in All Moss Genera with Known Gravitropic Protonemata

Moss protonemata from several species are known to be gravitropic. The characterization of additional gravitropic species would be valuable to identify conserved traits that may relate to the mechanism of gravitropism. In this study, four new species were found to have gravitropic protonemata, Fissidens adianthoides, Fissidens cristatus, Physcomitrium pyriforme, and Barbula unguiculata. Comparison of upright and inverted apical cells of P. pyriforme and Fissidens species showed clear axial sedimentation. This sedimentation is highly regulated and not solely dependent on amyloplast size. Additionally, the protonemal tip cells of these species contained broad subapical zones that displayed lateral amyloplast sedimentation. The conservation of a zone of lateral sedimentation in a total of nine gravitropic moss species from five different orders supports the idea that this sedimentation serves a specialized and conserved function in gravitropism, probably in gravity sensing.     

Effects of the myosin ATPase inhibitor 2,3-butanedione monoxime on amyloplast kinetics and gravitropism of Arabidopsis hypocotyls

The actin cytoskeleton is a crucial component in plant gravitropism, and studies confirm that alterations to actin filaments (F-actin) can have dramatic effects on gravitropic curvature in roots and shoots. Many models for gravisensing in higher plants suggest that the key to gravity perception and signal transduction lies in intimate interactions between F-actin and amyloplasts. In this study, we investigated gravitropism in hypocotyls by analyzing the effect of myosin inhibition on gravitropic curvature in order to clarify the role of the actomyosin system in shoot gravitropism. To study amyloplast movement in endodermal cells (i.e., gravity-perceiving statocytes) of living seedlings, we repositioned a confocal laser scanning microscope (CLSM) so that its rotatable stage was oriented vertically. Seedlings containing green fluorescent protein-labeled endodermal amyloplasts were incubated with the ATPase inhibitor 2,3-butanedione monoxime (BDM) and then mounted on the stage so that the hypocotyls were vertical. Using CLSM, we imaged the endodermal amyloplasts, while the hypocotyls were oriented vertically and also after they were reoriented by 90°. Our results show that BDM reduces gravitropic curvature in a concentration-dependent manner. In addition, BDM increases amyloplast movement in hypocotyls of vertical seedlings, but reduces amyloplast movement in hypocotyls of reoriented seedlings, suggesting that myosin may participate in the intracellular transport of amyloplasts in statocytes. These results can be explained in the context of amyloplasts as both noise indicators and gravity susceptors, with BDM producing less coherent amyloplast movement that results in an increased signal-to-noise ratio, which may account for at least part of the observed reduction in gravitopic curvature.     

Joining forces: The interface of gravitropism and plastid protein import

In flowering plants, gravity perception appears to involve the sedimentation of starch-filled plastids, called amyloplasts, within specialized cells (the statocytes) of shoots (endodermal cells) and roots (columella cells). Unfortunately, how the physical information derived from amyloplast sedimentation is converted into a biochemical signal that promotes organ gravitropic curvature remains largely unknown. Recent results suggest an involvement of the Translocon of the Outer Envelope of (Chloro) plastids (TOC) in early phases of gravity signal transduction within the statocytes. This review summarizes our current knowledge of the molecular mechanisms that govern gravity signal transduction in flowering plants and summarizes models that attempt to explain the contribution of TOC proteins in this important behavioral plant growth response to its mechanical environment.Key words: gravitropism, root, amyloplast, TOC complex, TOC132, TOC75     

Ethylene modulates flavonoid accumulation and gravitropic responses in roots of Arabidopsis

Plant organs change their growth direction in response to reorientation relative to the gravity vector. We explored the role of ethylene in Arabidopsis (Arabidopsis thaliana) root gravitropism. Treatment of wild-type Columbia seedlings with the ethylene precursor 1-aminocyclopropane carboxylic acid (ACC) reduced root elongation and gravitropic curvature. The ethylene-insensitive mutants ein2-5 and etr1-3 had wild-type root gravity responses, but lacked the growth and gravity inhibition by ACC found in the wild type. We examined the effect of ACC on tt4(2YY6) seedlings, which have a null mutation in the gene encoding chalcone synthase, the first enzyme in flavonoid synthesis. The tt4(2YY6) mutant makes no flavonoids, has elevated indole-3-acetic acid transport, and exhibits a delayed gravity response. Roots of tt4(2YY6), the backcrossed line tt4-2, and two other tt4 alleles had wild-type sensitivity to growth inhibition by ACC, whereas the root gravitropic curvature of these tt4 alleles was much less inhibited by ACC than wild-type roots, suggesting that ACC may reduce gravitropic curvature by altering flavonoid synthesis. ACC treatment induced flavonoid accumulation in root tips, as judged by a dye that becomes fluorescent upon binding flavonoids in wild type, but not in ein2-5 and etr1-3. ACC also prevented a transient peak in flavonoid synthesis in response to gravity. Together, these experiments suggest that elevated ethylene levels negatively regulate root gravitropism, using EIN2- and ETR1-dependent pathways, and that ACC inhibition of gravity response occurs through altering flavonoid synthesis.     

Negative gravitropic response of roots directs auxin flow to control root gravitropism

Root tip is capable of sensing and adjusting its growth direction in response to gravity, a phenomenon known as root gravitropism. Previously, we have shown that negative gravitropic response of roots (NGR) is essential for the positive gravitropic response of roots. Here, we show that NGR, a plasma membrane protein specifically expressed in root columella and lateral root cap cells, controls the positive root gravitropic response by regulating auxin efflux carrier localization in columella cells and the direction of lateral auxin flow in response to gravity. Pharmacological and genetic studies show that the negative root gravitropic response of the ngr mutants depends on polar auxin transport in the root elongation zone. Cell biology studies further demonstrate that polar localization of the auxin efflux carrier PIN3 in root columella cells and asymmetric lateral auxin flow in the root tip in response to gravistimulation is reversed in the atngr1;2;3 triple mutant. Furthermore, simultaneous mutations of three PIN genes expressed in root columella cells impaired the negative root gravitropic response of the atngr1;2;3 triple mutant. Our work revealed a critical role of NGR in root gravitropic response and provided an insight of the early events and molecular basis of the positive root gravitropism.     

Diversity of root hydrotropism among natural variants of Arabidopsis thaliana

Root gravitropism affects root hydrotropism. The interference intensity of root gravitropism with root hydrotropism differs among plant species. However, these differences have not been well compared within a single plant species. In this study, we compared root hydrotropism in various natural variants of Arabidopsis under stationary conditions. As a result, we detected a range of root hydrotropism under stationary conditions among natural Arabidopsis variants. Comparison of root gravitropism and root hydrotropism among several Arabidopsis natural variants classified natural variants that decreased root hydrotropism into two types; namely one type that expresses root gravitropism and root hydrotropism weaker than Col-0, and the other type that expresses weaker root hydrotropism than Col-0 but expresses similar root gravitropism with Col-0. However, root hydrotropism of all examined Arabidopsis natural variants was facilitated by clinorotation. These results suggested that the interference of root gravitropism with root hydrotropism is conserved among Arabidopsis natural variants, although the intensity of root gravitropism interference with root hydrotropism differs.

     

Role of endodermal cell vacuoles in shoot gravitropism

In higher plants, shoots and roots show negative and positive gravitropism, respectively. Data from surgical ablation experiments and analysis of starch deficient mutants have led to the suggestion that columella cells in the root cap function as gravity perception cells. On the other hand, endodermal cells are believed to be the statocytes (that is, gravity perceiving cells) of shoots. Statocytes in shoots and roots commonly contain amyloplasts which sediment under gravity. Through genetic research with Arabidopsis shoot gravitropism mutants, sgr1/scr and sgr7/shr, it was determined that endodermal cells are essential for shoot gravitropism. Moreover, some starch biosynthesis genes and EAL1 are important for the formation and maturation of amyloplasts in shoot endodermis. Thus, amyloplasts in the shoot endodermis would function as statoliths, just as in roots. The study of the sgr2 and zig/sgr4 mutants provides new insights into the early steps of shoot gravitropism, which still remains unclear. SGR2 and ZIG/SGR4 genes encode a phospholipase-like and a v-SNARE protein, respectively. Moreover, these genes are involved in vacuolar formation or function. Thus, the vacuole must play an important role in amyloplast sedimentation because the sgr2 and zig/sgr4 mutants display abnormal amyloplast sedimentation.     

An Arabidopsis E3 ligase, SHOOT GRAVITROPISM9, modulates the interaction between statoliths and F-actin in gravity sensing

Higher plants use the sedimentation of amyloplasts in statocytes as statolith to sense the direction of gravity during gravitropism. In Arabidopsis thaliana inflorescence stem statocyte, amyloplasts are in complex movement; some show jumping-like saltatory movement and some tend to sediment toward the gravity direction. Here, we report that a RING-type E3 ligase SHOOT GRAVITROPISM9 (SGR9) localized to amyloplasts modulates amyloplast dynamics. In the sgr9 mutant, which exhibits reduced gravitropism, amyloplasts did not sediment but exhibited increased saltatory movement. Amyloplasts sometimes formed a cluster that is abnormally entangled with actin filaments (AFs) in sgr9. By contrast, in the fiz1 mutant, an ACT8 semidominant mutant that induces fragmentation of AFs, amyloplasts, lost saltatory movement and sedimented with nearly statically. Both treatment with Latrunculin B, an inhibitor of AF polymerization, and the fiz1 mutation rescued the gravitropic defect of sgr9. In addition, fiz1 decreased saltatory movement and induced amyloplast sedimentation even in sgr9. Our results suggest that amyloplasts are in equilibrium between sedimentation and saltatory movement in wild-type endodermal cells. Furthermore, this equilibrium is the result of the interaction between amyloplasts and AFs modulated by the SGR9. SGR9 may promote detachment of amyloplasts from AFs, allowing the amyloplasts to sediment in the AFs-dependent equilibrium of amyloplast dynamics.