Piriformospora indica, an endophytic fungus of the order Sebacinales, interacts with the roots of a large variety of plant species. We compared the interaction of this fungus with Chinese cabbage ...(Brassica campestris subsp. chinensis) and Arabidopsis seedlings. The development of shoots and roots of Chinese cabbage seedlings was strongly promoted by P. indica and the fresh weight of the seedlings increased approximately twofold. The strong stimulation of root hair development resulted in a bushy root phenotype. The auxin level in the infected Chinese cabbage roots was twofold higher compared with the uncolonized controls. Three classes of auxin-related genes, which were upregulated by P. indica in Chinese cabbage roots, were isolated from a double-subtractive expressed sequence tag library: genes for proteins related to cell wall acidification, intercellular auxin transport carrier proteins such as AUX1, and auxin signal proteins. Overexpression of B. campestris BcAUX1 in Arabidopsis strongly promoted growth and biomass production of Arabidopsis seedlings and plants; the roots were highly branched but not bushy when compared with colonized Chinese cabbage roots. This suggests that BcAUX1 is a target of P. indica in Chinese cabbage. P. indica also promoted growth of Arabidopsis seedlings but the auxin levels were not higher and auxin genes were not upregulated, implying that auxin signaling is a more important target of P. indica in Chinese cabbage than in Arabidopsis. The fungus also stimulated growth of Arabidopsis aux1 and aux1/axr4 and rhd6 seedlings. Furthermore, a component in an exudate fraction from P. indica but not auxin stimulated growth of Chinese cabbage and Arabidopsis seedlings. We propose that activation of auxin biosynthesis and signaling in the roots might be the cause for the P. indica-mediated growth phenotype in Chinese cabbage.
Ca2+, a versatile intracellular second messenger in various signaling pathways, initiates many responses involved in growth, defense and tolerance to biotic and abiotic stress. Endogenous and ...exogenous signals induce cytoplasmic Ca2+ (Ca2+cyt) elevation, which are responsible for the appropriate downstream responses.
Here we report on an ethyl-methane sulfonate-mediated Arabidopsis mutant that fails to induce Ca2+cyt elevation in response to exudate preparations from the pathogenic mibrobes Alternaria brassicae, Rhizoctonia solani, Phytophthora parasitica var. nicotianae and Agrobacterium tumefaciens. The cytoplasmic Ca2+elevation mutant1 (cycam1) is susceptible to infections by A. brassicae, its toxin preparation and sensitive to abiotic stress such as drought and salt. It accumulates high levels of reactive oxygen species and contains elevated salicylic acid, abscisic acid and bioactive jasmonic acid iso-leucine levels. Reactive oxygen species- and phytohormone-related genes are higher in A. brassicae-treated wild-type and mutant seedlings. Depending on the analysed response, the elevated levels of defense-related compounds are either caused by the cycam mutation and are promoted by the pathogen, or they are mainly due to the pathogen infection or application of pathogen-associated molecular patterns. Furthermore, cycam1 shows altered responses to abscisic acid treatments: the hormone inhibits germination and growth of the mutant.
We isolated an Arabidopsis mutant which fails to induce Ca2+cyt elevation in response to exudate preparations from various microbes. The higher susceptibility of the mutant to pathogen infections correlates with the higher accumulation of defense-related compounds, such as phytohormones, reactive oxygen-species, defense-related mRNA levels and secondary metabolites. Therefore, CYCAM1 couples Ca2+cyt elevation to biotic, abiotic and oxidative stress responses.
Redox Responsive Transcription Factor1 (RRTF1) in Arabidopsis is rapidly and transiently upregulated by H202, as well as biotic- and abiotic-induced redox signals. RRTF1 is highly conserved in angio- ...sperms, but its physiological role remains elusive. Here we show that inactivation of RRTF1 restricts and overexpression promotes reactive oxygen species (ROS) accumulation in response to stress. Transgenic lines overexpressing RRTF1 are impaired in root and shoot development, light sensitive, and susceptible to Alternaria brassicae infection. These symptoms are diminished by the beneficial root endophyte Piriformospora indica, which reduces ROS accumulation locally in roots and systemi- cally in shoots, and by antioxidants and ROS inhibitors that scavenge ROS. More than 800 genes were detected in mature leaves and seedlings of transgenic lines overexpressing RRTF1; ∽40% of them have stress-, redox-, ROS-regulated-, ROS-scavenging-, defense-, cell death- and related functions. Bioinformatic analyses and in vitro DNA binding assays demonstrate that RRTF1 binds to GCC-box-like sequences in the promoter of RRTFl-responsive genes. Upregulation of RRTF1 by stress stimuli and H202 requires WRKY18/40/60. RRTF1 is co-regulated with the phylogenet- ically related RAP2.6, which contains a GCC-box-like sequence in its promoter, but transgenic lines overexpressing RAP2.6 do not accumulate higher ROS levels. RRTF1 also stimulates systemic ROS accumulation in distal non-stressed leaves. We conclude that the elevated levels of the highly conserved RRTF1 induce ROS accumulation in response to ROS and ROS-producing abiotic and biotic stress signals.
Phytoplasma suspected symptoms of little leaf, flat stem, witches’ broom and leaf yellowing were recorded on the four legume species, cowpea (
Vigna unguiculata
(L.) Walp.), pigeon pea (
Cajanus ...cajan
(L.) Millsp.), lentil (
Lens culinaris
Medikus) and mung bean (
Vigna radiata
(L.) Wilczek) in the states of Delhi, Uttar Pradesh (UP) and Kerala from 2014 to 2016. DNA specific fragments of approximately 1.3 kb were amplified from symptomatic samples of cowpea, pigeon pea, lentil and mung bean in nested PCR assays by using two sets of universal phytoplasma nested specific primers P1/P7 followed by 3Far/3Rev. No DNA amplifications were observed in any of the non-symptomatic legume samples with same primer pairs. Pair wise sequence comparison, phylogeny and virtual RFLP analysis of 16S rDNA sequences of the four legume species confirmed the association of four different groups and subgroups of phytoplasmas in the present study. The mung bean witches’ broom at Delhi was identified to be associated with strain related to ‘
Ca.
P. aurantifolia’ (16SrII-D), pigeon pea little leaf at Faizabad, UP with strain related to ‘
Ca.
P. phoenicium’ (16SrIX-C), lentil witches’ broom at Faizabad, UP with ‘
Ca. P. trifolii
’ (16SrVI-D) and cow pea flat stem disease at Kerala with ‘
Ca.
P. cynodontis’ (16SrXIV-A). Association of ‘
Ca
. P. cynodontis' (16SrXIV-A) infecting cowpea, ‘
Ca. P. trifolii
’ (16SrVI-D) in lentil and phytoplasmas strain related to ‘
Ca.
P. phoenicium’ (16SrIX-C) infecting pigeon pea are the new reports to the world.
The growth-promoting and root-colonizing endophyte Piriformospora indica induces camalexin and the expression of CYP79B2, CYP79B3, CYP71A13, PAD3, and WRKY33 required for the synthesis of ...indole-3-acetaldoxime (IAOx)-derived compounds in the roots of Arabidopsis seedlings. Upregulation of the mRNA levels by P. indica requires cytoplasmic calcium elevation and mitogen-activated protein kinase 3 but not root-hair-deficient 2, radical oxygen production, or the 3-phosphoinositide-dependent kinase 1/oxidative signal-inducible 1 pathway. Because P. indica-mediated growth promotion is impaired in cyp79B2 cyp79B3 seedlings, while pad3 seedlings-which do not accumulate camalexin-still respond to the fungus, IAOx-derived compounds other than camalexin (e.g., indole glucosinolates) are required during early phases of the beneficial interaction. The roots of cyp79B2 cyp79B3 seedlings are more colonized than wild-type roots, and upregulation of the defense genes pathogenesis-related (PR)-1, PR-3, PDF1.2, phenylalanine ammonia lyase, and germin indicates that the mutant responds to the lack of IAOx-derived compounds by activating other defense processes. After 6 weeks on soil, defense genes are no longer upregulated in wild-type, cyp79B2 cyp79B3, and pad3 roots. This results in uncontrolled fungal growth in the mutant roots and reduced performance of the mutants. We propose that a long-term harmony between the two symbionts requires restriction of root colonization by IAOx-derived compounds.
During a survey performed in sapota orchards of India, from 2015 to 2018, symptoms of phyllody, little leaf, flat stem and witches’ broom were observed in three states: Karnataka, Kerala and Tripura. ...The association of phytoplasmas was confirmed in all the symptomatic sapota samples by using nested PCR specific primers (P1/P7, R16F2n/R16R2 and 3Far/3Rev) with amplification of fragments of ~ 1.25 kb and ~ 1.3 kb. Association of three phytoplasma groups, aster yellows with flat stem from Tripura (Lembucherra), clover proliferation with phyllody symptoms at Karnataka (Bengaluru) and bermuda grass white leaf with flat stem and little leaf from Kerala (Thiruvananthapuram) and Tripura (Cocotilla) were confirmed by
16S rRNA
gene sequence comparison analysis. Virtual RFLP analysis of
16S rRNA
gene sequences using pDRAW32 further classified the sapota phytoplasma isolates into 16SrI-B, 16SrVI-D and 16SrXIV-A subgroups. This is the first report on identification of three phytoplasma groups in sapota in world.
Soil is a non-renewable natural resource which forms all ecosystems on the earth and provides the basis for food production for heterotrophic organisms, including men. The increase in the world ...population requires also an increase in agricultural production which was and is mainly achieved by massive use of mineral nutrients. However, the experience of the last century has demonstrated that the high mineral input has severe consequences for the ecosystems. An alternative more environmentally friendly strategy for agricultural production is provided by the nature itself: Beneficial root-colonizing fungi and bacteria have tremendous impact on the performance of agricultural plants (crops). Understanding of these symbioses requires knowledge about the communication between the partners. The microbes often release bioactive compounds into the rhizosphere which activate signaling or transport processes and thus promote plant performance. A general new concept for fertilizers in the agriculture could be to utilize microbe-derived bio-effectors in combination with appropriate nutrient supplies to promote biomass and yield production of agricultural plants while simultaneously reducing the input of agrochemicals. Here, we describe some concepts for the identification and utilization of microbial preparations and microbe-derived bio-effectors for the improvement of the performance of agricultural plants, using the root-colonizing endophytic fungus Piriformospora indica.