Mutualistic association of Paecilomyces formosus LHL10 offers thermotolerance to Cucumis sativus

We investigated in this study the influence of an endophytic fungus, Paecilomyces formosus LHL10, on the thermotolerance of cucumber (Cucumis sativus) upon exposure to high (38°C) and low (8°C) temperature stresses. The results showed that endophyte-inoculated plants had significantly higher plant growth attributes under high-temperature stress. However, they were either low or insignificant in non-inoculated control and inoculated plants with 8°C treatments. Lower stress-promulgated water deficit and cellular membrane damage were observed in endophyte-treated plants after 38°C treatment than in control plants under 8°C stress. Total polyphenol, reduced glutathione, and lipid peroxidation activities were reduced in endophyte-associated plants after exposure to 38°C as compared with control and 8°C-treated plants. The concentration of saturated fatty acids (palmitic-C16:0; stearic-C18:0) was lower in endophyte-treated plants with or without low-temperature stress, but after 8°C treatment increased compared with controls. Unsaturated fatty acids (oleic-C18:1; linoleic-C18:2; linolenic-C18:3 acids) were similar at normal conditions; however, at 38°C, C18:2 and C18:3 were decreased, and C18:1 was increased in endophyte-treated plants compared with controls, while the inverse relationship was found at 8°C. Low levels of abscisic acid in P. formosus-associated plants after 38°C treatments revealed stress tolerance compared with control and 8°C-treated plants. In contrast, salicylic acid was pronounced in endophyte-treated plants after low-temperature stress as compared to other treatments. The results provide evidence that the response to P. formosus association was beneficial at normal growth temperature and had varying effects in response to temperature stress.     

Endophytic Cephalotheca sulfurea AGH07 reprograms soybean to higher growth

Abstract

Gibberellins (GAs) are well known for plant growth promotion. GAs production by fungi has received little attention, although substantial work has been carried out on other aspects of plant growth-promoting fungi (PGPF). We investigated GAs production and plant growth-promoting capacity of an endophytic fungus isolated from the roots of soil grown soybean plants. The endophytic fungus is reported as GAs producer and as PGPF for the first time in this study. Nine endophytic isolates were collected from the roots of soybean, and culture filtrates (CFs) obtained from their pure cultures were screened on Waito-C, a dwarf rice cultivar, for the presence of GAs. Of these, seven fungal isolates promoted shoot length as compared to control (distilled water), while one inhibited it. Three fungal isolates were selected on the basis of higher shoot elongation as compared to wild type Gibberella fujikuroi, which was used as positive control. The growth-prompting capacity of selected fungal isolates SB5-1, SB3-2, and SB3-3 was bio-assayed on soybean cv. Hwangkeumkong. Fungal isolate SB5-1 provided maximum plant height (31.6 cm), shoot length (21.1 cm), whole plant fresh biomass (2.41 g), shoot fresh biomass (1.99 g), and leaf area (24.37 cm²). The CF of isolate SB5-1 was analyzed for the presence of GAs, and it was found that all physiologically active GAs were present (GA₁, 0.15 ng/ml, GA₃, 1.2 ng/ml, GA₄, 7.37 ng/ml, and GA₇, 3.18 ng/ml) in conjunction with physiologically inactive GA₅, GA₉, GA₁₅, GA₁₉, GA₂₀, and GA₂₄. The fungal isolate SB5-1 was identified as a new strain of Cephalotheca sulfurea through molecular and phylogenetic approaches.     

miRNAs: Major modulators for crop growth and development under abiotic stresses

Cumulatively, biotic and abiotic stresses of various magnitudes can decrease the production of crops by 70%. miRNAs have emerged as a genetic tool with enormous potential that can be exploited to understand stress tolerance at the molecular level and eventually regulate stress in crops. Plant miRNA targets frequently fit into diverse families of TFs that control the expression of genes related to a certain trait. As key machinery in gene regulatory networks, it is agreed that a broad understanding of miRNAs will greatly increase our understanding of plant responses to environmental stresses. miRNA-led stress regulatory networks are being considered as novel tools for the development of abiotic stress tolerance in crops. At this time, we need to expand our knowledge about the modulatory role of miRNAs during environmental fluctuations. It has become exceedingly clear that with increased understanding of the role of miRNAs during stress, the techniques for using miRNA-mediated gene regulation to enhance plant stress tolerance will become more effective and reliable. In this review we present: (1) miRNAs as a potential avenue for the modulation of abiotic stresses, and (2) summarize the research progress regarding plant responses to stress. Current progress is explained through discussion of the identification and validation of several miRNAs that enhance crop tolerance of salinity, drought, etc., while missing links on different aspects of miRNAs related to abiotic stress tolerance are noted.     

Pure culture of <Emphasis Type="Italic">Metarhizium anisopliae</Emphasis> LHL07 reprograms soybean to higher growth and mitigates salt stress

Little is known about the role of endophytic fungi against abiotic stresses and isoflavonoids (IF) contents of soybean. In current study, we investigated the role of fungal endophytes on the growth of soybean under salt stress conditions. Pure cultures of nine endophytic fungi were isolated from the roots of field-grown soybean plants, and their culture filtrates were screened on Waito-C and Dongjin-byeo rice cultivars; for identification of plant growth promoting fungal strains. It was observed that fungal isolate GMC-2B significantly promoted the growth of both Waito-C and Dongjin-byeo. GMC-2B was later identified as a new strain of Metarhizium anisopliae LHL07 on the basis of 18S rDNA sequences and phylogenetic analysis. Metarhizium anisopliae LHL07 inoculated soybean plants recorded significantly higher shoot length, shoot fresh and dry biomass, chlorophyll contents, transpiration rate, photosynthetic rate and leaf area; under sodium chloride induced salt stress as compared to non-inoculated control plants. An elevated proline and reduced superoxide dismutase and malondialdehyde contents in M. anisopliae LHL07 inoculated soybean plants demonstrated mitigation of salt induced oxidative stress. Furthermore, reduced abscisic acid and elevated jasmonic acid contents in soybean plants confirmed that lesser stress was convened to M. anisopliae inoculated-plants under salinity stress. We also assessed the role of M. anisopliae interaction on IF biosynthesis of soybean, and found significantly higher IF contents in M. anisopliae inoculated soybean plants. In conclusion, endophytic fungal interactions with soybean can be beneficial to improve soybean quality and quantity under salt affected agricultural systems.     

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.     

Agrobacterium cucumeris sp. nov. isolated from crazy roots on cucumber (Cucumis sativus)

Three plant rhizogenic strains O132^T, O115 and O34 isolated from Cucumis sp. L. were assessed for taxonomic affiliation by using polyphasic taxonomic methods. Based on the results of the sequence analysis of the 16S rRNA and multilocus sequence analysis (MLSA) of the three housekeeping genes atpD, recA and rpoB, all the strains were clustered within the genus Agrobacterium where they form a novel branch. Their closest relative was Agrobacterium tomkonis (genomospecies G3). Moreover, digital DNA-DNA hybridization (dDDH) and average nucleotide identity (ANI) comparisons between strains O132^T and O34 and their closest relatives provided evidence that they constitute a new species, because the obtained values were significantly below the threshold considered as a borderline for the species delineation. Whole-genome phylogenomic analysis also indicated that the cucumber strains are located within the separate, well-delineated biovar 1 sub-clade of the genus Agrobacterium. Furthermore, the physiological and biochemical properties of these strains allowed to distinguish them from their closest related species of the genus Agrobacterium. As a result of the performed overall characterization, we propose a new species as Agrobacterium cucumeris sp. nov., with O132^T (=CFBP 8997^T = LMG 32451^T) as the type strain.     

Water relations and photosynthesis in Cucumis sativus L. leaves under salt stress

Hydroponically grown cucumber plants were exposed to 14-d period of salinity (0, 50, 100 mM NaCl). NaCl caused reduction in the relative water content in the leaves. The Na⁺ content increased and the K⁺ content decreased. The net photosynthetic rate, stomatal conductance and transpiration rate were markedly decreased by all of the salt treatments. Salinity decreased also the maximum quantum efficiency of photosystem 2 (PS 2) determined as the variable to maximum fluorescence ratio, the photochemical quantum yield of PS 2 and the photochemical fluorescence quenching, while the non-photochemical quenching increased. Above results indicate that NaCl affects photosynthesis through both stomata closure and non-stomatal factors.     

Gibberellin production and plant growth promotion from pure cultures of Cladosporium sp. MH-6 isolated from cucumber (Cucumis sativus L.)

Gibberellin (GA) production by soil fungi has received little attention, although substantial work has been carried out on other aspects of plant growth promoting fungi (PGPF). In our studies we investigated GA production and growth-promoting capacity of a novel fungal strain isolated from the roots of soil-grown cucumber. Pure cultures of 19 endophytic fungi were tested for shoot length promotion of Waito-C rice to identify the GA production capacity of these fungal isolates. Isolate MH-6 significantly increased shoot length (12.9 cm) of Waito-C, in comparison to control treatments. Bioassay with culture filtrate (CF) of MH-6 also significantly promoted growth attributes of cucumber plants. Analysis of MH-6 CF showed the presence of physiologically active (GA1, 1.97 ng/mL; GA3, 5.18 ng/mL; GA4, 13.35 ng/mL and GA7, 2.4 ng/ mL) in conjunction with physiologically inactive (GA9 [0.69 ng/mL], GA12 [0.24 ng/mL], GA15 [0.68 ng/ mL, GA19 [1.94 ng/mL and GA20 [0.78 ng/mL]) gibberellins. The CF of MH-6 produced greater amounts of GA3, GA4, GA7 and GA19 than wild type Fusarium fujikuroi, a fungus known for high production of GA. The fungal isolate MH-6 was identified as a new strain of Cladosporium sp. on the basis of sequence homology (99%) and phylogenetic analysis of 18S rDNA sequence.     

外源一氧化氮对盐胁迫下黄瓜幼苗生长和渗透调节物质含量的影响

采用水培法,研究了外源一氧化氮(NO)对黄瓜幼苗生长和渗透调节物质含量的影响。结果表明:正常生长条件下添加外源NO能促进黄瓜幼苗生长,而添加NO信号传递途径关键酶——鸟苷酸环化酶抑制剂亚甲基蓝(MB-1)显著抑制了黄瓜幼苗的生长;盐胁迫条件下,添加外源NO明显缓解了盐胁迫对黄瓜幼苗生长的抑制,与单独盐处理比较,株高、茎粗、鲜质量、干质量显著增加,渗透调节物质如可溶性糖、可溶性蛋白和脯氨酸含量明显提高,而MB-1能够不同程度地消除NO的这些调节作用;NO对盐胁迫下黄瓜幼苗生长影响大于正常生长条件下黄瓜幼苗,NO的作用可能是通过鸟苷酸环化酶介导的。     

The role of nitric oxide in plant growth regulation and responses to abiotic stresses

Nitric oxide (NO) has received much attention in the recent two decades, equally from human, animal and plant biologists. It was found to play a crucial role in human and animal physiology, immunological reactions and signal transduction. Its ubiquity and versatile properties caught the attention of plant physiologists and biochemists. This work presents an extensive review on the NO presence and action in plants. Various modes of NO synthesis are discussed and the most novel approaches to the elucidation of plant nitric oxide synthase (NOS) structure are presented. This review focuses on the physiological role of NO in regulation of plant growth and development, as well as in the process of gene expression. Special attention is given to the action of NO during abiotic stress and the antioxidant properties of the molecule.     

外源亚精胺对根际低氧胁迫下黄瓜幼苗光合作用的影响

采用营养液水培法,研究了低氧胁迫下亚精胺(Spd)对黄瓜幼苗净光合速率（P_n）、细胞间隙CO₂浓度（C_i）、气孔导度（G_s）、蒸腾速率（T_r）以及量子效率（Φ_c）和羧化效率（CE）的影响.结果表明,低氧胁迫下,黄瓜植株P_n呈下降趋势,处理10 d后达最低值,为同期对照的63.33%,而低氧胁迫的外源Spd处理10 d时P_n升高了1.25倍;C_i与P_n呈一定负相关性（R²=0.4730～0.7118）,G_s与T_r的变化幅度较大,低氧胁迫下有明显下降趋势,Spd处理后其值有所上升,两者呈显著相关（R²=0.7821～0.9458）,但与P_n的相关性不显著;低氧下Φ_c和CE比对照分别下降了63.01%和72.33%,而Spd处理后,Φ_c和CE值分别提高了23%和14%.表明在低氧胁迫下黄瓜幼苗的光合抑制主要是由非气孔限制所引起的,而外源Spd可通过对光系统的修饰减轻黄瓜幼苗的低氧胁迫伤害.     

Growth and development of cucumber Cucumis sativus L. in the prereproductive period under long photoperiods

When plants are grown in a greenhouse or in controlled environment growth rooms, prolonging the photoperiod, including towards continuous light, is one of the ways to increase plant productivity and energy savings. However, exposing some plant species to long photoperiods causes leaf injuries and growth reductions. We studied the effect of the photoperiod (8, 12, 16, 20, and 24 h) and photosynthetic photon flux density (60, 120, and 160 μmol/m² s PAR) on cucumber plants Cucumis sativus L. in a prereproductive period. It was shown that the response of the cucumber plants to a photoperiod over 20 h, including continuous light, depending on the plant age and light intensity, may include leaf movement or paraheliotropism, non-photochemical energy dissipation, and/or reversible photoinhibition of a reaction center of photosystem II, development of reversible chlorosis, reduction of a light-harvesting complex, and increase in the content of carotenoids. Reaction of immature and virginile plants to long photoperiods was different, which high-lights the need for experimental separation of the prereproductive period in terms of age states and consideration of this when developing crop production plan.     

不同生态环境下雌雄同株黄瓜单性结实性遗传的比较

以两个单性结实性不同的雌雄同株黄瓜自交系构建的4世代群体为试材,采用植物数量性状主基因+多基因混合遗传模型联合分析方法,对南京江宁和河北昌黎两地雌雄同株黄瓜单性结实性遗传进行了比较研究.结果表明：不同生态环境下,雌雄同株黄瓜单性结实性遗传均符合E-1-1模型,受2对加性-显性-上位性主基因+加性-显性多基因控制,存在基因型与环境互作效应.但是,不同环境条件下F₁的遗传倾向和遗传参数不同,F₂的主基因遗传率为42.1%～97.5%,其遗传差异主要由黄瓜结果期两地日照和温度差异所致.强单性结实黄瓜品种选育以双亲均为强单性结实材料为宜,杂种后代宜在不同生态条件下选择鉴定.     

Interaction between arbuscular mycorrhizal fungus Glomus mosseae and plant growth promoting fungus Phoma sp. on their root colonization and growth promotion of cucumber (Cucumis sativus L.)

Interaction between arbuscular mycorrhizal fungus Glomus mosseae and plant growth promoting fungus Phoma sp. was studied for its effect on their root colonization and plant growth of cucumber. Two isolates of Phoma sp. (GS8-2 and GS8-3) were tested with G. mosseae. The percent root length colonized by G. mosseae was not adversely affected by the presence of Phoma isolates. In contrast, the root colonization of both isolates GS8-2 and GS8-3 in 4-week-old plants was significantly reduced (80.7% and 84.3%, respectively) by added G. mosseae. Inoculating plants with each Phoma isolate significantly increased the shoot dry weight. However, dual inoculation of each Phoma isolate with G. mosseae had no significant effect on growth enhancement.     

Ultrastructure of root cup meristematic cells of Cucumis sativus L. seedlings under inhibition of root growth by synthetic cyclopentene beta,beta'-triketone

A study was made of the ultrastructure of root cap meristematic cells in the seedlings of Cucumis sativus L., whose growth was inhibited by the action of 2-acetyl-4-metylcyclopent-4-en-1,3-dione (AMCD). It is shown that the treatment of seeds by this substance at a concentration of 100 microg/ml leads to the main root inhibition by 73% and, as compared to the control, causes the highest changes in mitochondria that do not reach normal dimensions and undergo degeneration. AMCD caused significant changes in the cell walls that in test samples unusually became electron-dense. Deposits of osmiophylic substance, presumably consisting of terpenoid-like compounds, were often found in the cytoplasm of treated cells as well as in the extracellular space. In many cells, plasmalemma and amiloplast membranes underwent destructive changes. We discuss a possible relation between inhibitory effect of AMCD on the root growth, and its influence on the ultrastructure in investigated cells.     

Sugar signalling and gene expression in relation to carbohydrate metabolism under abiotic stresses in plants

Sucrose is required for plant growth and development. The sugar status of plant cells is sensed by sensor proteins. The signal generated by signal transduction cascades, which could involve mitogen-activated protein kinases, protein phosphatases, Ca²⁺ and calmodulins, results in appropriate gene expression. A variety of genes are either induced or repressed depending upon the status of soluble sugars. Abiotic stresses to plants result in major alterations in sugar status and hence affect the expression of various genes by down- and up-regulating their expression. Hexokinase-dependent and hexokinase-independent pathways are involved in sugar sensing. Sucrose also acts as a signal molecule as it affects the activity of a proton-sucrose symporter. The sucrose transporter acts as a sucrose sensor and is involved in phloem loading. Fructokinase may represent an additional sensor that bypasses hexokinase phosphorylation especially when sucrose synthase is dominant. Mutants isolated on the basis of response of germination and seedling growth to sugars and reporter-based screening protocols are being used to study the response of altered sugar status on gene expression. Commoncis-acting elements in sugar signalling pathways have been identified. Transgenic plants with elevated levels of sugars/sugar alcohols like fructans, raffinose series oligosaccharides, trehalose and mannitol are tolerant to different stresses but have usually impaired growth. Efforts need to be made to have transgenic plants in which abiotic stress responsive genes are expressed only at the time of adverse environmental conditions instead of being constitutively synthesized.