寄生植物对寄主植物的化学识别

植物间寄生关系的研究近年来受到了广泛的重视。大量的研究表明,寄主释放的次生物质对植物间寄生关系的建立和维持起了重要的调节作用。寄主植物的次生物质对寄生植物的化学防御和昆虫授粉等生态功能起重要的作用,寄主植物次生物质对寄生植物生理与生态的调节作用是受寄生植物基因调节的。更为重要的是寄主植物释放的次生物质成为寄生植物的种子萌发和吸器发生的异源识别物质。能够刺激寄生植物种子萌发的次生物质主要是倍半萜和氢醌类物质,而诱导吸器发生的物质则是酚酸、醌和黄酮类化合物,诱导吸器发生的核心结构是对苯醌。这些异源识别物质大多是寄主植物释放的化感抑制物质,显示寄生植物在化学防御方面要比寄主植物高级。异源识别化合物的活性与其氧化潜力显著相关。由于寄生植物中存在一抑制异源识别物质诱导吸器发生的调节过程,因此吸器的产生与寄生植物根部接触异源识别物质的浓度与时间呈正相关关系,这一调节过程对寄生植物准确识别寄主并寄生其上是十分重要的。对寄生植物和寄主植物间的化学识别关系的揭示有助于人们防治有害寄生植物和开发利用有价值的寄生植物资源。     

氮磷供给对两种马先蒿根系形态建成的影响

采用溶液培养法在无寄主伴生的条件下对三色马先蒿（Pedicularis tricolor）和大王马先蒿（P. rex）进行培养,分别分析了不同氮磷供给水平（标准Long Ashton营养液、缺氮、缺磷、2倍氮、2倍磷）及不同氮素形态（硝态氮和铵态氮）对两种马先蒿根系形态建成的影响。结果表明：1）氮磷供给水平对两种马先蒿的主根长、侧根长、侧根数量以及吸器发生的影响存在种间差异,缺氮和缺磷均能促进三色马先蒿主根和侧根的伸长生长及吸器的产生,而相同条件下大王马先蒿根系生长受到抑制,且没有吸器产生;2）氮素形态对两种马先蒿根系形态建成及生长发育存在影响：1倍硝态氮的处理中,三色马先蒿的根系生长最好,生物量积累最多,相同条件下大王马先蒿根系生长和生物量积累与标准营养液处理无差异,而仅供应铵态氮对二者的生长不利。此外,发现三色马先蒿的吸器产生不需要机械刺激。初步推测,两种马先蒿的吸器产生机制或进程存在差异,氮和磷可能参与了吸器发生过程的调控。     

云南哀牢山两种常见半寄生植物的生态化学计量特征及其与寄主的关系

为深入探究半寄生植物与其寄主间的养分关系,在云南哀牢山徐家坝地区选取两种常见半寄生植物椆树桑寄生(Loranthus delavayi)和柳叶钝果寄生(Taxillus delavayi),研究其寄主枝条-吸器-寄生枝条-寄生叶片间的碳(C)、氮(N)、磷(P)生态化学计量特征关系。结果表明:1)两种半寄生植物的寄主枝条-吸器-寄生枝条-寄生叶片这一连续体的C、N、P生态化学计量特征变化趋势并不完全相同,具有物种自身的特性。2)同一半寄生植物的寄主枝条具有相似的C、N、P生态化学计量特征,寄主物种对半寄生植物的生态化学计量特征没有显著影响。3)寄主枝条的C、N、P生态化学计量特征间具有紧密的相关关系,吸器弱于寄主枝条,寄生枝条弱于吸器,寄生叶片的N、P含量相关关系不显著。4)寄主枝条与寄生叶片间的C含量存在极显著负相关关系。5)吸器与寄主枝条间的C、N、P生态化学计量特征存在紧密的相关关系,但在生态化学计量特征的数值上吸器与寄生枝条更为接近。吸器作为连接寄主与寄生植物的关键部位,其与寄主枝条极为显著的相关关系体现了寄主枝条养分对寄生植物的重要性,而吸器在元素含量、计量比的数值以及相互关系上与寄生枝条更为相似,则体现了吸器作为寄生植物器官具有与寄生枝条相似的生理功能。通过对寄主枝条-吸器-寄生枝条-寄生叶片C、N、P生态化学计量特征的分析,为深入研究半寄生植物的养分利用策略与生态适应性提供了重要的基础资料。     

云南哀牢山两种常见半寄生植物的生态化学计量特征及其与寄主的关系

为深入探究半寄生植物与其寄主间的养分关系, 在云南哀牢山徐家坝地区选取两种常见半寄生植物椆树桑寄生(Loranthus delavayi)和柳叶钝果寄生(Taxillus delavayi), 研究其寄主枝条-吸器-寄生枝条-寄生叶片间的碳(C)、氮(N)、磷(P)生态化学计量特征关系。结果表明: 1)两种半寄生植物的寄主枝条-吸器-寄生枝条-寄生叶片这一连续体的C、N、P生态化学计量特征变化趋势并不完全相同, 具有物种自身的特性。2)同一半寄生植物的寄主枝条具有相似的C、N、P生态化学计量特征, 寄主物种对半寄生植物的生态化学计量特征没有显著影响。3)寄主枝条的C、N、P生态化学计量特征间具有紧密的相关关系, 吸器弱于寄主枝条, 寄生枝条弱于吸器, 寄生叶片的N、P含量相关关系不显著。4)寄主枝条与寄生叶片间的C含量存在极显著负相关关系。5)吸器与寄主枝条间的C、N、P生态化学计量特征存在紧密的相关关系, 但在生态化学计量特征的数值上吸器与寄生枝条更为接近。吸器作为连接寄主与寄生植物的关键部位, 其与寄主枝条极为显著的相关关系体现了寄主枝条养分对寄生植物的重要性, 而吸器在元素含量、计量比的数值以及相互关系上与寄生枝条更为相似, 则体现了吸器作为寄生植物器官具有与寄生枝条相似的生理功能。通过对寄主枝条-吸器-寄生枝条-寄生叶片C、N、P生态化学计量特征的分析, 为深入研究半寄生植物的养分利用策略与生态适应性提供了重要的基础资料。     

四种马先蒿属植物的菌根形态学研究

通过4种马先蒿属植物,三色马先蒿三色变种(PedicularistricolorHand.-Mazz.var.tricolor),管花马先蒿管花变种(P.siphonanthaDonvar.siphonantha),全缘叶马先蒿全缘亚种(P.integrifoliaHk.f.subsp.integerrimaTsoong)和二歧马先蒿(P.ditrotomaBonati)根的解剖,在这四种马先蒿属植物根内均发现有真菌菌丝侵入,真菌菌丝向外伸入土壤,向内侵入皮层细胞内形成囊泡.从根表面分离到的真菌厚垣孢子,经鉴定均为漏斗球囊霉(GlomusmosseaeNicol.&Gerd.).其中,三色马先蒿和二歧马先蒿栽培实验表明,真菌起到了改善寄主植物营养状况的作用.     

青藏高原“黑土滩”常见毒草对甘肃马先蒿的化感作用

以青藏高原"黑土滩"典型毒草黄花棘豆(Oxytropis ochrocephala)、瑞香狼毒(Stellera chamaejasme)、黄帚橐吾(Ligularia virgaurea)、密花香薷(Elsholtzia densa)和南山蒿(Artemisia nanschanica)为供体材料,在实验室条件下测试了它们的根、茎、叶水浸提液对甘肃马先蒿(Pedicularis kansuensis)的化感作用,以期寻找对甘肃马先蒿种子萌发和幼苗生长具有生态化感抑制作用的植物种类及部位。结果表明,随着5种毒草各器官水浸提液浓度的升高,其对甘肃马先蒿化感作用增强;且同一植物在测试浓度相同的条件下,表现为叶、根的水浸提液对甘肃马先蒿种子和幼苗的抑制作用强于茎的浸提液。从5种毒草化感作用的综合表现看,黄花棘豆的叶和根、南山蒿的叶在浸提液浓度为25 g/L时对甘肃马先蒿幼苗生长的抑制作用显著大于其他毒草;当作用浓度升至100 g/L时,对甘肃马先蒿种子萌发和幼苗生长都表现出致死作用。因此,对黄花棘豆叶、根和南山蒿叶等植物器官的化感作用应作进一步的研究,将有望开发出针对甘肃马先蒿的专用生物源控制剂。     

尕海湿地不同密度下甘肃马先蒿根系分叉数与连接数、分支角度的关系

根系构型决定根系分布及其对养分和水分的吸收效率,是植物根系与胁迫生境相互适应的结果。采用标准化主轴估计(Standardized major axis estimation, SMA)法,并采用全根挖掘和Win-RHIZO根系分析仪相结合的方法,按照甘肃马先蒿种群密度设置I(10—31株/m~2)、Ⅱ(32—53株/m~2)、Ⅲ(54—75株/m~2)3个样地,研究了尕海湿地不同密度下甘肃马先蒿(Pedicularis kansuensis)根系分叉数与连接数及分支角度的关系。结果表明:随着种群密度由高到低转变,甘肃马先蒿的高度、盖度、地上生物量、根系分叉数及外部连接数逐渐减小,地下生物量、根系内部连接数、分支角度逐渐增大;甘肃马先蒿根系分叉数与内部连接数、外部连接数及分支角度均呈异速生长关系,随着种群密度由高到低转变,甘肃马先蒿根系内部连接数与分支角度增加的速度逐渐大于分叉数与外部连接数减小的速度,根系分叉数与内部连接数的异速斜率逐渐减小,与外部连接数、分支角度的异速斜率逐渐增大。甘肃马先蒿在高密度倾向于密集型根系构型构建模式,在低密度选择扩散型根系生长模式,体现了高寒湿地植物种群应对资源多重竞争的生态适应机制。     

檀香小G蛋白Rac1基因的克隆与功能分析

植物Rac蛋白属于小分子G蛋白ROP家族,广泛参与活性氧(ROS)产生、激素信号转导和组织形态建成.檀香(Santalum album Linn.)是著名的珍贵树种,为半寄生植物,其正常生长需要根部特化的吸器从其他寄主植物摄取营养物质.该研究基于全长转录组数据,采用RT-PCR方法克隆得到了1个檀香Rac基因,命名为S...     

肉苁蓉寄生生长形态发育

肉苁蓉(Cistanche deserticola)是一种沙生根寄生濒危药用植物。利用光镜和电镜手段, 采用人工培养和诱导等方法详细观察并研究了肉苁蓉寄生生长过程中种子萌发、吸器产生以及植物体形态发育的过程。结果表明: (1)人工可以诱导肉苁蓉的种子萌发。肉苁蓉种胚具有明显的极性, 珠孔端细胞小于合点端, 珠孔端细胞分化、生长并产生白色的类胚根状结构。(2)有些化学物质可以诱导初生吸器的产生。用2, 6二甲氧基-对苯醌培养肉苁蓉24-26小时后, 其先端膨大并产生突起, 形成类似根毛状的结构, 即初生吸器。(3) 肉苁蓉属于主动寄生植物, 其在初生吸器与寄主幼根黏连后产生次生吸器。肉苁蓉与寄主梭梭(Haloxylon ammodendron)共培养, 其初生吸器主动与寄主幼根(0.1 mm左右)黏连, 穿过寄主根表皮和皮层后与维管束连通形成次生吸器, 肉苁蓉植物体分化、发育的基本组织形成。被寄生后的寄主根横向发育加快, 同时肉苁蓉植物体开始分化和发育。(4)肉苁蓉可以寄生在寄主幼根(0.1 mm左右)的任意部位。     

寄生植物的生物学特性及生态学效应

寄生植物是陆地生态系统的重要组成部分,在多数生态系统中均有分布.寄生植物完全或部分依赖于寄主植物获取有机养分、无机养分和水分等,以满足自身生长发育需求.在生态学效应方面,寄生植物不但直接影响寄主植物的生长发育,也会通过改变寄主植物的竞争力而影响植物群落组成.寄生植物对寄主植物个体水平的影响多是消极的,并能给人类的生产活动造成直接经济损失,故对寄生植物的防治一直是人们关注的热点问题;在群落水平上,寄生植物对促进植物群落的物种多样性有积极的意义,基于此,寄生植物有潜在利用价值.深入了解寄生植物的生理生态学特性,对有害寄生植物的防除和有益寄生植物的利用有重要价值.本文对寄生植物的生物学特性,包括其寄生类型、寄主范围及偏好、寄主识别和吸器的形成与功能,进行了阐述,并就寄生植物对寄主植物和群落结构的影响进行了探讨,对相关研究前景进行了展望.     

In Vitro Haustorium Development in Roots and Root Cultures of the Hemiparasitic Plant Triphysaria Versicolor

Parasitic plants in the Orobanchaceae invade host plant roots through root organs called haustoria. Parasite roots initiate haustorium development when exposed to specific secondary metabolites that are released into the rhizosphere by host plant roots. While molecular approaches are increasingly being taken to understand the genetic mechanism underlying these events, a limitation has been the lack of a transformation system for parasitic plants. Since the haustorium development occurs in roots of Orobanchaceae, root cultures may be suitable material for transient or stable transformation experiments. To this end, root cultures were obtained from explants, and subsequently calluses, from the hemiparasitic plant Triphysaria versicolor. The cultured roots retained their competence to form haustoria when exposed to host roots, host root exudates, or purified haustorium-inducing factors. The root culture haustoria invaded host roots and initiated a vascular continuity between the parasite and host roots. The ontogeny of haustoria development on root cultures was indistinguishable from that on seedlings roots. Root cultures should provide useful material for molecular studies of haustorium development.     

Three-dimensional reconstructions of haustoria in two parasitic plant species in the Orobanchaceae

Parasitic plants infect other plants by forming haustoria, specialized multicellular organs consisting of several cell types, each of which has unique morphological features and physiological roles associated with parasitism. Understanding the spatial organization of cell types is, therefore, of great importance in elucidating the functions of haustoria. Here, we report a three-dimensional (3-D) reconstruction of haustoria from two Orobanchaceae species, the obligate parasite Striga hermonthica infecting rice (Oryza sativa) and the facultative parasite Phtheirospermum japonicum infecting Arabidopsis (Arabidopsis thaliana). In addition, field-emission scanning electron microscopy observation revealed the presence of various cell types in haustoria. Our images reveal the spatial arrangements of multiple cell types inside haustoria and their interaction with host roots. The 3-D internal structures of haustoria highlight differences between the two parasites, particularly at the xylem connection site with the host. Our study provides cellular and structural insights into haustoria of S. hermonthica and P. japonicum and lays the foundation for understanding haustorium function.

Three-dimensional image reconstruction visualized the spatial organization of cell types in the haustoria of the Orobanchaceae parasitic plants Striga hermonthica and Phtheirospermum japonicum.     

Fast and abundant in vitro spontaneous haustorium formation in root hemiparasitic plant Pedicularis kansuensis Maxim. (Orobanchaceae)

Haustorium formation is the characteristic feature of all parasitic plants and a vital process for successful parasitism. Previous investigations on haustorium initiation and development are constricted to induced processes by host-derived signals or synthetic analogs. Spontaneous haustorium formation in the absence of host signals, a process representing an early stage in the evolution of parasitic plants, remains largely unexplored. Lack of fast and frequent formation of spontaneous haustoria greatly hinders full understanding of haustorium formation in root hemiparasites. In this study, seedlings of Pedicularis kansuensis Maxim., a facultative root hemiparasitic species in Orobanchaceae observed to produce many spontaneous haustoria, were grown in autoclaved water agar in the absence of any known haustoriuminducing stimulants. We aimed to test the temporal and developmental pattern of spontaneous haustorium formation. Also, effects of sucrose supply and root contact on spontaneous haustorium formation were tested. Spontaneous haustoria were observed starting from six days after germination, much earlier than previously reported root hemiparasites. A majority of the spontaneous haustoria formed on lateral roots. Percentage of seedlings with spontaneous haustoria was 28.8% when grown on water agar plates, with a mean of four haustoria per seedling two weeks after germination. Haustorium formation by seedlings grown in water agar amended with 2% sucrose was more than twice of those without sucrose amendment. Singly grown seedlings were able to develop spontaneous haustoria at similar levels as those grown with another conspecific seedling. In view of the fast and abundant formation of spontaneous haustoria, P. kansuensis may be developed as an excellent experimental system in future investigations for unraveling endogenous regulation of haustorium initiation and development in root hemiparasitic plants.     

Inoculation with arbuscular mycorrhizal fungi suppresses initiation of haustoria in the root hemiparasite Pedicularis tricolor

Background and Aims

Plant parasitism and arbuscular mycorrhizal (AM) associations have many parallels and share a number of regulatory pathways. Despite a rapid increase in investigations addressing the roles of AM fungi in regulating interactions between parasitic plants and their hosts, few studies have tested the effect of AM fungi on the initiation and differentiation of haustoria, the parasite-specific structures exclusively responsible for host attachment and nutrient transfer. In this study, we tested the influence of AM fungi on haustorium formation in a root hemiparasitic plant.

Methods

Using a facultative root hemiparasitic species (Pedicularis tricolor) with the potential to form AM associations, the effects of inoculation were tested with two AM fungal species, Glomus mosseae and Glomus intraradices, on haustorium initiation in P. tricolor grown alone or with Hordeum vulgare ‘Fleet’ (barley) as the host plant. This study consisted of two greenhouse pot experiments.

Key Results

Both AM fungal species dramatically suppressed intraspecific haustorium initiation in P. tricolor at a very low colonization level. The suppression over-rode inductive effects of the parasite''s host plant on haustoria production and caused significant growth depression of P. tricolor.

Conclusions

AM fungi had strong and direct suppressive effects on haustorium formation in the root hemiparasite. The significant role of AM fungi in haustorium initiation of parasitic plants was demonstrated for the first time. This study provides new clues for the regulation of haustorium formation and a route to development of new biocontrol strategies in management of parasitic weeds.     

Subtilase activity in intrusive cells mediates haustorium maturation in parasitic plants

Parasitic plants that infect crops are devastating to agriculture throughout the world. These parasites develop a unique inducible organ called the haustorium that connects the vascular systems of the parasite and host to establish a flow of water and nutrients. Upon contact with the host, the haustorial epidermal cells at the interface with the host differentiate into specific cells called intrusive cells that grow endophytically toward the host vasculature. Following this, some of the intrusive cells re-differentiate to form a xylem bridge (XB) that connects the vasculatures of the parasite and host. Despite the prominent role of intrusive cells in host infection, the molecular mechanisms mediating parasitism in the intrusive cells remain poorly understood. In this study, we investigated differential gene expression in the intrusive cells of the facultative parasite Phtheirospermum japonicum in the family Orobanchaceae by RNA-sequencing of laser-microdissected haustoria. We then used promoter analyses to identify genes that are specifically induced in intrusive cells, and promoter fusions with genes encoding fluorescent proteins to develop intrusive cell-specific markers. Four of the identified intrusive cell-specific genes encode subtilisin-like serine proteases (SBTs), whose biological functions in parasitic plants are unknown. Expression of SBT inhibitors in intrusive cells inhibited both intrusive cell and XB development and reduced auxin response levels adjacent to the area of XB development. Therefore, we propose that subtilase activity plays an important role in haustorium development in P. japonicum.

Subtilases specifically expressed in intrusive cells regulate auxin-mediated host–parasite connections in the parasitic plant Phtheirospermum japonicum.     

Agrobacterium rhizogenes-mediated transformation of the parasitic plant Phtheirospermum japonicum

Background

Plants within the Orobanchaceae are an agriculturally important group of parasites that attack economically important crops to obtain water and nutrients from their hosts. Despite their agricultural importance, molecular mechanisms of the parasitism are poorly understood.

Methodology/Principal Findings

We developed transient and stable transformation systems for Phtheirospermum japonicum, a facultative parasitic plant in the Orobanchaceae. The transformation protocol was established by a combination of sonication and acetosyringone treatments using the hairy-root-inducing bacterium, Agrobacterium rhizogenes and young seedlings. Transgenic hairy roots of P. japonicum were obtained from cotyledons 2 to 3 weeks after A. rhizogenes inoculation. The presence and the expression of transgenes in P. japonicum were verified by genomic PCR, Southern blot and RT-PCR methods. Transgenic roots derived from A. rhizogenes-mediated transformation were able to develop haustoria on rice and maize roots. Transgenic roots also formed apparently competent haustoria in response to 2,6-dimethoxy-1,4-benzoquinone (DMBQ), a haustorium-inducing chemical. Using this system, we introduced a reporter gene with a Cyclin B1 promoter into P. japonicum, and visualized cell division during haustorium formation.

Conclusions

We provide an easy and efficient method for hairy-root transformation of P. japonicum. Transgenic marker analysis revealed that cell divisions during haustorium development occur 24 h after DMBQ treatment. The protocols described here will allow functional analysis of genes involved in plant parasitism.     

Arbuscular mycorrhizal fungi: potential biocontrol agents against the damaging root hemiparasite Pedicularis kansuensis?

Spatial expansion of root hemiparasitic Pedicularis kansuensis in Bayanbulak Grassland of Xinjiang Uygur Autonomous Region (China) has caused great loss of herbage yield and has threatened the local livestock industry. Current management practices using manual eradication and chemical control have been proved problematic. Arbuscular mycorrhizal (AM) fungi have been suggested to be potential biocontrol agents against a number of plant pests, but experimental evidence is lacking against weedy P. kansuensis. In this study, we tested the hypothesis that inoculation with AM fungi will cause growth depression in P. kansuensis and reduce its damage to host plants. Based on the confirmation of AM status and host community of the hemiparasite in the field, a pot cultivation experiment was conducted to test the influence of an AM fungus (Glomus mosseae) on growth of P. kansuensis and the parasitized host (Elymus nutans). AM colonization was observed in roots of P. kansuensis, but the levels were much lower than those of its adjacent host species. A negative correlation between AM levels and the numbers of haustoria was detected for the field samples of the hemiparasite. Strong suppression of haustorium formation, a significant reduction in plant dry weight (DW), as well as marked reduction in the survival rate of P. kansuensis after inoculation with AM fungi was observed. In contrast, inoculation with G. mosseae increased root DW and whole plant DW of parasitized host plants. Our findings demonstrated significantly repressive effects of AM fungi on growth performance of P. kansuensis with and without the presence of a host. The potential of AM fungi as biocontrol agents against the damaging hemiparasite was confirmed.