The pepper extracellular peroxidase CaPO2 is required for salt, drought and oxidative stress tolerance as well as resistance to fungal pathogens

In plants, biotic and abiotic stresses regulate the expression and activity of various peroxidase isoforms. Capsicum annuum EXTRACELLULAR PEROXIDASE 2 (CaPO2) was previously shown to play a role in local and systemic reactive oxygen species bursts and disease resistance during bacterial pathogen infection. Here, we report CaPO2 expression patterns and functions during conditions of biotic and abiotic stress. In pepper plants, CaPO2 expression was strongly induced by abscisic acid, but not by defense-related plant hormones such as salicylic acid, ethylene and jasmonic acid. CaPO2 was also strongly induced by abiotic and biotic stress treatments, including drought, cold, high salinity and infection by the hemibiotrophic fungal pathogen Colletotrichum coccodes. Loss-of-function of CaPO2 in virus-induced gene silenced pepper plants led to increased susceptibility to salt- and osmotic-induced stress. In contrast, CaPO2 overexpression in transgenic Arabidopsis thaliana plants conferred enhanced tolerance to high salt, drought, and oxidative stress, while also enhancing resistance to infection by the necrotrophic fungal pathogen Alternaria brassicicola. Taken together, these results provide evidence for the involvement of pepper extracellular peroxidase CaPO2 in plant defense responses to various abiotic stresses and plant fungal pathogens.     

Microbial rescue to plant under habitat-imposed abiotic and biotic stresses

Habitat-imposed abiotic and biotic stress is a serious condition and is also a land-degradation problem in arid and semi-arid regions, causing major problem for crop productivity. Most of the cultivable and a least half of irrigated lands around the world are severely affected by environmental stresses. However, in these conditions, there are plant populations successfully adapted and evolutionarily different in their strategy of stress tolerance. Vascular plants do not function as autonomous individuals, but house diverse communities of symbiotic microbes. The role of these microbes can no longer be ignored. Microbial interactions are critical not only for host but also for fungal survival in stressed environments. Plants benefit extensively by harboring these associated microbes; they promote plant growth and confer enhanced resistance to various pathogens by producing antibiotics. To date, improvements in plant quality, production, abiotic and biotic stress resistance, nutrient, and water use have relied largely on manipulating plant genomes by breeding and genetic modification. Increasing evidence indicates that the function of symbiotic microbes seems to parallel more than one of these characteristics.     

Retrospect and prospects of plant metabolic engineering

With the advancement of biotechnological tools and techniques such as next generation sequencing, RNAomics, epigenomics, gene silencing, plant, microbe transformation, proteomics and metabolomics, the understanding of metabolic pathways and their manipulation for the desired characters became feasible. Metabolic engineering has been successful in the production of golden rice, bioprocess for artemisinin production, flavonoids in plant and microbes as well as generated biotic and abiotic stress tolerance in several crop plants. In view of the significance of metabolic engineering, this article includes recent techniques developed and their use in manipulation of glyoxalase metabolism for multiple abiotic stress tolerance in plants. The importance of engineering of flavonoids pathway for high value antioxidants production as well as improving the biotic and abiotic stress tolerance has been documented. Importance and success of metabolic engineering has been realized by its promising hope for sustainable technologies of bioactives production for mankind’s health as well as in the generation of improved crop varieties.

     

Unraveling the role of fungal symbionts in plant abiotic stress tolerance

Fungal symbionts have been found to be associated with every plant studied in the natural ecosystem, where they colonize and reside entirely or partially in the internal tissues of their host plant. Fungal endophytes can express/form a range of different lifestyle/relationships with different host including symbiotic, mutualistic, commensalistic and parasitic in response to host genotype and environmental factors. In mutualistic association fungal endophyte can enhance growth, increase reproductive success and confer biotic and abiotic stress tolerance to its host plant. Since abiotic stress such as, drought, high soil salinity, heat, cold, oxidative stress and heavy metal toxicity is the common adverse environmental conditions that affect and limit crop productivity worldwide. It may be a promising alternative strategy to exploit fungal endophytes to overcome the limitations to crop production brought by abiotic stress. There is an increasing interest in developing the potential biotechnological applications of fungal endophytes for improving plant stress tolerance and sustainable production of food crops. Here we have described the fungal symbioses, fungal symbionts and their role in abiotic stress tolerance. A putative mechanism of stress tolerance by symbionts has also been covered.Key words: abiotic stress, endophytes, fungal symbiont, mycorrhizal fungus, Piriformospora indica, stress tolerance, symbiosis     

The Comparative Analysis of Osmotins and Osmotin-Like PR-5 Proteins

Abstract: One of the ways that plants respond to biotic and/or abiotic stress factors is the accumulation of pathogenesis-related proteins of class 5 (PR-5), which are evolutionary conserved in the plant kingdom. Within the PR-5 family, a distinct subgroup of osmotin and closely related proteins has been characterized. In contrast to the extracellular forms of PR-5 proteins, osmotins presumably accumulate in the vacuole of the cell. They contain a C-terminal propeptide that is considered to be a determinant for vacuolar targeting. The comparison of the three-dimensional structure of tobacco PR-5 d with the sequences of some osmotins showed that the proteins consist of three conserved domains, with the acidic cleft between domains I and II. Besides the constitutive species and tissue-specific presence, the osmotins are also induced by several abiotic and biotic stresses. Among them, fungal infections can elicit osmotin gene expression, and most known proteins from the family have antifungal activity in in vitro assays. In agreement with the osmotin structure and data on the activity of similar proteins, a two-step mechanism, which involves reaction of osmotins with the fungal wall and the permeabilization of fungal membranes, is discussed.     

Role of microorganisms in adaptation of agriculture crops to abiotic stresses

Increased incidences of abiotic and biotic stresses impacting productivity in principal crops are being witnessed all over the world. Extreme events like prolonged droughts, intense rains and flooding, heat waves and frost damages are likely to further increase in future due to climate change. A wide range of adaptations and mitigation strategies are required to cope with such impacts. Efficient resource management and crop/livestock improvement for evolving better breeds can help to overcome abiotic stresses to some extent. However, such strategies being long drawn and cost intensive, there is a need to develop simple and low cost biological methods for the management of abiotic stress, which can be used on short term basis. Microorganisms could play a significant role in this respect, if we can exploit their unique properties of tolerance to extremities, their ubiquity, genetic diversity, their interaction with crop plants and develop methods for their successful deployment in agriculture production. Besides influencing the physico-chemical properties of rhizospheric soil through production of exopolysaccharides and formation of biofilm, microorganisms can also influence higher plants response to abiotic stresses like drought, chilling injury, salinity, metal toxicity and high temperature, through different mechanisms like induction of osmo-protectants and heat shock proteins etc. in plant cells. Use of these microorganisms per se can alleviate stresses in crop plants thus opening a new and emerging application in agriculture. These microbes also provide excellent models for understanding the stress tolerance, adaptation and response mechanisms that can be subsequently engineered into crop plants to cope with climate change induced stresses.     

Expression of apoplastically secreted tobacco osmotin in cotton confers drought tolerance

Osmotin or osmotin-like proteins have been shown to be induced in several plant species in response to various types of biotic and abiotic challenges. The protein is generally believed to be involved in protecting the plant against these stresses. Although some understanding of the possible mechanism underlying the defense function of osmotin against biotic stresses is beginning to emerge, its role in abiotic stress response is far from clear. We have transformed cotton plants with a tobacco-osmotin gene, lacking the sequence encoding its 20 amino acid-long, C-terminal vacuolar-sorting motif, under the control of CaMV 35S promoter. Apoplastic secretion of the recombinant protein was confirmed and the plants were evaluated for their ability to tolerate drought conditions. Under polyethylene glycol-mediated water stress, the osmotin-expressing seedlings showed better growth performance. The transformants showed a slower rate of wilting during drought and faster recovery following the termination of dry conditions in a greenhouse setting. During drought, the leaves from transgenic plants had higher relative water content and proline levels, while showing reduced H₂O₂ levels, lipid peroxidation and electrolyte leakage. Importantly, following a series of dry periods, the osmotin transformants performed better in terms of most growth and developmental parameters tested. Most relevant, the fiber yield of transgenic plants did not suffer as much as that of their non-transgenic counterparts under drought conditions. The results provide direct support for a protective role of osmotin in cotton plants experiencing water stress and suggest a possible way to achieve tolerance to drought conditions by means of genetic engineering. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Fungal growth promotor endophytes: a pragmatic approach towards sustainable food and agriculture

Agricultural productivity suffers a heavy loss due to plant pathogens, insect pests and various abiotic stresses. Agriculture being the world’s largest economic sector, it is the need of time to find and establish the ideal strategy for sustainable agriculture and improvement in crop growth. Endophytes are microorganisms that asymptomatically grow within the plant tissues without causing any disease to the host. Endophytic fungi live in symbiotic association with plants and play an important role in plant growth promotion, higher seed yield and plants resistant to various biotic, abiotic stresses and diseases. Many are able to produce antimicrobial compounds, plant growth hormones and various agrochemical bioactive metabolites. These mycoendophytes hold enormous potential for the development of eco-friendly and economically viable agricultural products. In this review we focused on the endophytic fungi recovered from different medicinal plants, their active principles involved in plant growth enhancement and the applications of fungal endophytes in agriculture. Moreover, we also discussed about endophytic fungi and their pragmatic approach towards sustainable food and agriculture.     

Overexpression of a tobacco osmotin gene in carrot (Daucus carota L.) enhances drought tolerance

Osmotin and osmotin-like proteins belong to the PR-5 pathogenesis-related group of proteins and are induced in response to various types of biotic and abiotic stresses in several plant species. Carrot was transformed with a tobacco osmotin gene that encodes a protein lacking the vacuolar-sorting motif that is composed of a 20-amino-acid sequence at the C-terminal end, under the control of the cauliflower mosaic virus 35S promoter, using Agrobacterium-mediated transformation. Transgene integration and expression were confirmed by Southern and western blot analyses, and three selected transgenic lines were evaluated for their ability to tolerate drought stress. Under drought stress conditions, all transformants exhibited slower rates of wilting compared with the wild-type plants and recovered faster when the drought stress was alleviated. Transformants showed lower levels of hydrogen peroxide accumulation, reduced lipid peroxidation and electrolyte leakage, and higher leaf water content under drought stress. Our results provide additional evidence for the protective ability of the osmotin protein against drought stress conditions and suggest a possible means to achieve tolerance against this abiotic stress in plants.     

Expression of Arabidopsis Bax Inhibitor‐1 in transgenic sugarcane confers drought tolerance

The sustainability of global crop production is critically dependent on improving tolerance of crop plants to various types of environmental stress. Thus, identification of genes that confer stress tolerance in crops has become a top priority especially in view of expected changes in global climatic patterns. Drought stress is one of the abiotic stresses that can result in dramatic loss of crop productivity. In this work, we show that transgenic expression of a highly conserved cell death suppressor, Bax Inhibitor‐1 from Arabidopsis thaliana (AtBI‐1), can confer increased tolerance of sugarcane plants to long‐term (>20 days) water stress conditions. This robust trait is correlated with an increased tolerance of the transgenic sugarcane plants, especially in the roots, to induction of endoplasmic reticulum (ER) stress by the protein glycosylation inhibitor tunicamycin. Our findings suggest that suppression of ER stress in C₄ grasses, which include important crops such as sorghum and maize, can be an effective means of conferring improved tolerance to long‐term water deficit. This result could potentially lead to improved resilience and yield of major crops in the world.     

Trichoderma/pathogen/plant interaction in pre-harvest food security

Large losses before crop harvesting are caused by plant pathogens, such as viruses, bacteria, oomycetes, fungi, and nematodes. Among these, fungi are the major cause of losses in agriculture worldwide. Plant pathogens are still controlled through application of agrochemicals, causing human disease and impacting environmental and food security. Biological control provides a safe alternative for the control of fungal plant pathogens, because of the ability of biocontrol agents to establish in the ecosystem. Some Trichoderma spp. are considered potential agents in the control of fungal plant diseases. They can interact directly with roots, increasing plant growth, resistance to diseases, and tolerance to abiotic stress. Furthermore, Trichoderma can directly kill fungal plant pathogens by antibiosis, as well as via mycoparasitism strategies. In this review, we will discuss the interactions between Trichoderma/fungal pathogens/plants during the pre-harvest of crops. In addition, we will highlight how these interactions can influence crop production and food security. Finally, we will describe the future of crop production using antimicrobial peptides, plants carrying pathogen-derived resistance, and plantibodies.     

Role of Pathogen-Related Protein 10 (PR 10) under Abiotic and Biotic Stresses in Plants

Members of the Pathogenesis Related (PR) 10 protein family have been identified in a variety of plant species and a wide range of functions ranging from defense to growth and development has been attributed to them. PR10 protein possesses ribonuclease (RNase) activity, interacts with phytohormones, involved in hormone-mediated signalling, afforded protection against various phytopathogenic fungi, bacteria, and viruses particularly in response to biotic and abiotic stresses. The resistance mechanism of PR10 protein may include activation of defense signalling pathways through possible interacting proteins involved in mediating responses to pathogens, degradation of RNA of the invading pathogens. Moreover, several morphological changes have been shown to accompany the enhanced abiotic stress tolerance. In this review, the possible mechanism of action of PR10 protein against biotic and abiotic stress has been discussed. Furthermore, our findings also confirmed that the in vivo Nitric oxide (NO) is essential for most of environmental abiotic stresses and disease resistance against pathogen infection. The proper level of NO may be necessary and beneficial, not only in plant response to the environmental abiotic stress, but also to biotic stress. The updated information on this interesting group of proteins will be useful in future research to develop multiple stress tolerance in plants.     

High-efficiency transformation and selective tolerance against biotic and abiotic stress in mulberry, <Emphasis Type="Italic">Morus indica</Emphasis> cv. K2, by constitutive and inducible expression of tobacco <Emphasis Type="Italic">osmotin</Emphasis>

Osmotin and osmotin-like proteins are stress proteins belonging to the plant PR-5 group of proteins induced in several plant species in response to various types of biotic and abiotic stresses. We report here the overexpression of tobacco osmotin in transgenic mulberry plants under the control of a constitutive promoter (CaMV 35S) as well as a stress-inducible rd29A promoter. Southern analysis of the transgenic plants revealed the stable integration of the introduced genes in the transformants. Real-time PCR analysis provided evidence for the expression of osmotin in the transgenic plants under both the constitutive and stress-inducible promoters. Transgenic plants with the stress-inducible promoter were observed to better tolerate salt and drought stress than those with the constitutive promoter. Transgenic plants when subjected to simulated salinity and drought stress conditions showed better cellular membrane stability (CMS) and photosynthetic yield than non-transgenic plants under conditions of both salinity and drought stress. Proline levels were very high in transgenic plants with the constitutive promoter relative to those with the stress-inducible promoter. Fungal challenge undertaken with three fungal species known to cause serious losses to mulberry cultivation, namely, Fusarium pallidoroseum, Colletotrichum gloeosporioides and Colletotrichum dematium, revealed that transgenic plants with osmotin under control of the constitutive promoter had a better resistance than those with osmotin under the control of the stress-inducible promoter. Evaluation in next generation was undertaken by studying bud break in transgenic and non-transgenic plants under simulated drought (2% polyethylene glycol) and salt stress (200 mM NaCl) conditions. The axillary buds of the selected transgenic lines had a better bud break percentage under stressed conditions than buds from non-transgenic mulberry lines. A biotic assay with Bombyx mori indicated that osmotin protein had no undesirable effect on silkworm rearing and feeding. We therefore conclude that 35S transgenic plants are better suited for both abiotic stress also biotic challenges (fungal), while the rd29A transgenic plants are more responsive to drought.     

Functional characterization of germin and germin-like protein genes in various plant species using transgenic approaches

The important role of germins (GER) and genes coding for germin-like proteins (GLP) in responses against various stresses in both homologous and heterologous systems is well validated. This review summarizes the work on their functional validation using various biotechnological approaches. The genes are widely expressed during a specific period of plant growth and development, and exhibit a pattern of evolutionary subfunctionalization at both the intracellular and whole plant level. Their applications against various biotic and abiotic stresses, especially against fungal pathogens, are enormous. Although the validation of these proteins against various stresses has led to the development of commercially and agronomically important transgenic plants, much work is still needed to exploit this ever-expanding repertoire of genes and deploy them for commercial use. Historical progress of genetic engineering in GERs and GLPs is reviewed, and future prospects for their potential role in crop improvement are highlighted.     

Developing stress tolerant plants through in vitro selection—An overview of the recent progress

Biotic and abiotic stresses impose a major threat to agriculture. Therefore, the efforts to develop stress-tolerant plants are of immense importance to increase crop productivity. In recent years, tissue culture based in vitro selection has emerged as a feasible and cost-effective tool for developing stress-tolerant plants. Plants tolerant to both the biotic and the abiotic stresses can be acquired by applying the selecting agents such as NaCl (for salt tolerance), PEG or mannitol (for drought tolerance) and pathogen culture filtrate, phytotoxin or pathogen itself (for disease resistance) in the culture media. Only the explants capable of sustaining such environments survive in the long run and are selected. In vitro selection is based on the induction of genetic variation among cells, tissues and/or organs in cultured and regenerated plants. The selection of somaclonal variations appearing in the regenerated plants may be genetically stable and useful in crop improvement. This review focuses on the progress made towards the development of stress-tolerant lines through tissue culture based in vitro selection. Plants have evolved many biochemical and molecular mechanisms to survive under stress conditions. The mechanisms of ROS (reaction oxygen species) generation and removal in plants under biotic and abiotic stress conditions have also been reviewed.