Raheem Shahzad

Endophytic infection alleviates biotic stress in sunflower through regulation of defence hormones, antioxidants and functional amino acids

Eco-friendly strategies pertinent to disease resistance in crops are desired for sustainable agriculture as they minimize chemical input into the agriculture industry. Fungal endophytes with phytohormones secreting capacity are well known for their plant growth promotion but have never been reported previously for their favourable role in plant resistance against pathogens. We studied the role of two fungal endophytes in promoting growth of sunflower (Helianthus annuus L.) and regulating hormone signalling networks involved in plant defence against fungal infection caused by Alternaria alternata. The two endophytic fungi (Penicillium citrinum LWL4 and Aspergillus terreus LWL5) promoted growth of sunflower and produced gibberellins (GA1, GA3, GA4, GA9, GA12, and GA20), organic acids (malic, quinic and succinic acid) and siderophore. In A. alternata infected plants, the sole or combined treatment of P. citrinum and A. terreus markedly reduced disease incidence and reprogrammed the host plants to higher growth. The signalling of plant-defence hormones, such as salicylic acid and jasmonic acid fluctuated with the application of sole or combined endophytes in the diseased plants. The endophyte-association regulated oxidative stress responses by activating glutathione and polyphenol oxidase, and down-regulating catalase and peroxidase. In diseased plants, the endophytic-symbiosis rescued plant growth by the overwintering synthesis of free amino acids. In conclusion, the inoculation of endophytes reprogrammed plant growth during disease incidence by regulating the defence-linked responses of the host plant. Exploring endophytic symbiosis can help achieve sustainable agriculture in an eco-friendly way without using fungicides.

Endophytic fungi promote plant growth and mitigate the adverse effects of stem rot: an example of Penicillium citrinum and Aspergillus terreus

Disease resistance is a highly desirable crop trait in the sustainable agricultural industry. Endophytic fungi with gibberellins-secreting potential are now widely known for their ability to stimulate plant growth, but their role in promoting disease resistance in plants has rarely been reported. We have studied the role of Penicillium citrinum LWL4 and Aspergillus terreus LWL5 in time-dependent manner on sunflower (Helianthus annuus L.) growth, disease resistance and their capacity for the regulation of hormone signaling networks involved in plant defense against the stem rot caused by Sclerotium rolfsii for 3, 6 and 12 days after treatment (DAT). Our results show that plant growth characteristics (i.e. shoot length, shoot diameter, shoot fresh/dry weight, transpiration, stomatal conductance, photosynthesis and chlorophyll content) were promoted in fungi-treated plants with or without the disease caused by Sclerotium rolfsii as compared to their respective controls in 3, 6 and 12 DAT. The negative impacts of stem rot in endophyte-treated diseased plants were greatly reduced in comparison to control diseased plants shown by low disease severity in 3, 6 and 12 DAT. Similarly, fungal endophytes in diseased plants relieved the biotic stress in time-dependent manner (3, 6 and 12 DAT) as shown by low level of endogenous salicylic acid and jasmonic acid contents and were significantly higher in control diseased plants. Furthermore, we observed that the Penicillium citrinum LWL4 association had a greater positive effect on sunflower plants than Aspergillus terreus LWL5. It was concluded that inoculation with fungal endophytes reprogramed plant growth during disease incidence by regulating responses associated with host plant defense. Management strategies involving endophytic symbiosis can help achieve sustainability in agriculture in an eco-friendly manner by reducing excessive fungicide use.

Bacterial endophytes from arid land plants regulate endogenous hormone content and promote growth in crop plants: an example of Sphingomonas sp. and Serratia marcescens

ABSTRACT The objective of the present study was to determine the potential plant growth-promoting action of bacterial endophytes isolated from arid land-dwelling plants under normal conditions. Overall, five bacterial endophytes LK11 (Sphingomonas sp. LK11), TP5 (Bacillus subtilis), MPB5.3 (B. subtilis subsp. Subtilis), S9 (B. subtilis subsp. Subtilis), and TP1 (Serratia marcescens) were evaluated based on morphological characteristics after isolation and purification. Phytohormonal analysis of these endophytes predicted indole acetic acid (IAA) production 12.31 ± 0.45 , 6.8 ± 0.59, and 10.5 ± 1.02 μM/mL in the culture broths of LK11, MPB5.3, and TP1, respectively. Under controlled greenhouse conditions, these endophytes were inoculated into soybean, and their growth-promoting characteristics were compared with those of non-phytohormone-producing endophytes. In terms of plant growth promotion, among IAA-producing endophytes, LK11 and TP1 greatly improved physiological characteristics such as shoot/root length, fresh/dry weight, and chlorophyll contents. However, the non-phytohormone-producing endophytes TP5 and S9 did not show a growth-promoting effect. Based on these results, plants inoculated with LK11 and TP1 along with a control were subjected to endogenous hormonal analysis and showed a significant increase in abscisic acid (457.30–398.55 vs. 205.93 ng/g D.W.) and a decrease in jasmonic acid content (50.07–85.07 vs. 93.90 ng/g D.W.), respectively. Total gibberellin content was found to significantly increase in endophyte-inoculated plants (155.43–146.94 ng/g D.W.) as compared to that in controls (113.76 ng/g D.W.). In summary, bacterial endophytes might be used to enhance crop plant physiological characteristics isolated from arid land-inhabiting plants under normal conditions.

The Complete Chloroplast Genome of Wild Rice (Oryza minuta) and Its Comparison to Related Species

Oryza minuta, a tetraploid wild relative of cultivated rice (family Poaceae), possesses a BBCC genome and contains genes that confer resistance to bacterial blight (BB) and white-backed (WBPH) and brown (BPH) plant hoppers. Based on the importance of this wild species, this study aimed to understand the phylogenetic relationships of O. minuta with other Oryza species through an in-depth analysis of the composition and diversity of the chloroplast (cp) genome. The analysis revealed a cp genome size of 135,094 bp with a typical quadripartite structure and consisting of a pair of inverted repeats separated by small and large single copies, 139 representative genes, and 419 randomly distributed microsatellites. The genomic organization, gene order, GC content and codon usage are similar to those of typical angiosperm cp genomes. Approximately 30 forward, 28 tandem and 20 palindromic repeats were detected in the O. minuta cp genome. Comparison of the complete O. minuta cp genome with another eleven Oryza species showed a high degree of sequence similarity and relatively high divergence of intergenic spacers. Phylogenetic analyses were conducted based on the complete genome sequence, 65 shared genes and matK gene showed same topologies and O. minuta forms a single clade with parental O. punctata. Thus, the complete O. minuta cp genome provides interesting insights and valuable information that can be used to identify related species and reconstruct its phylogeny.

Bacillus aryabhattai SRB02 tolerates oxidative and nitrosative stress and promotes the growth of soybean by modulating the production of phytohormones

Plant growth promoting rhizobacteria (PGPR) are diverse, naturally occurring bacteria that establish a close association with plant roots and promote the growth and immunity of plants. Established mechanisms involved in PGPR-mediated plant growth promotion include regulation of phytohormones, improved nutrient availability, and antagonistic effects on plant pathogens. In this study, we isolated a bacterium from the rhizospheric soil of a soybean field in Chungcheong buk-do, South Korea. Using 16S rRNA sequencing, the bacterium was identified as Bacillus aryabhattai strain SRB02. Here we show that this strain significantly promotes the growth of soybean. Gas chromatography—mass spectrometry analysis showed that SRB02 produced significant amounts of abscisic acid, indole acetic acid, cytokinin and different gibberellic acids in culture. SRB02-treated soybean plants showed significantly better heat stress tolerance than did untreated plants. These plants also produced consistent levels of ABA under heat stress and exhibited ABA-mediated stomatal closure. High levels of IAA, JA, GA12, GA4, and GA7, were recorded in SRB02-treated plants. These plants produced longer roots and shoots than those of control plants. B. aryabhattai SRB02 was found to be highly tolerant to oxidative stress induced by H2O2 and MV potentiated by high catalase (CAT) and superoxide dismutase (SOD) activities. SRB02 also tolerated high nitrosative stress induced by the nitric oxide donors GSNO and CysNO. Because of these attributes, B. aryabhattai SRB02 may prove to be a valuable resource for incorporation in biofertilizers and other soil amendments that seek to improve crop productivity.

Kinetin modulates physio-hormonal attributes and isoflavone contents of Soybean grown under salinity stress

Crop productivity continues to decline due to a wide array of biotic and abiotic stresses. Salinity is one of the worst abiotic stresses, as it causes huge losses to crop yield each year. Kinetin (Kn) has been reported as plant growth regulator since long, but its role in improving plant growth and food quality under saline conditions through mediating phytohormonal cross-talk is poorly studied. Current study was designed to evaluate the impact of exogenously applied Kn on growth, isoflovones and endogenous phytohormones of soybean grown under NaCl induced salt stress. Soybean plants were grown in perlite (semi hydroponic), and under controlled green-house conditions. Elevated levels of exogenous Kn significantly mitigated the adverse effect of NaCl and rescued plant growth attributes, i.e., plant height, fresh and dry biomass of soybean plants grown in all treatments. Higher diadzen, glycitin, and genistin contents were observed in plants treated with elevated Kn in the presence or absence of NaCl induce salt stress. The gibberellins (GAs) biosynthesis pathway was up-regulated by Kn as the bioactive GA1 and GA4 contents were significantly higher in Kn treated plants, as compared to control, while GAs level reduced in NaCl treated plants. Contrary to GAs, the abscisic acid contents declined with Kn but promoted in NaCl stressed soybean plants. The endogenous jasmonic acid and salicylic acid contents of soybean enhanced with elevated Kn application, but they showed an antagonistic response under salt stress. Current study supports the active role of Kn to ameliorate the adverse effects of salt stress on the growth and food quality of soybean. The favorable role of Kn toward soybean growth under salt stress may be attributed to its potential to modulate cross-talk between the various phytohormones involved in soybean growth and its resistance to salinity stress.

Plant growth-promoting endophytic bacteria versus pathogenic infections: an example of Bacillus amyloliquefaciens RWL-1 and Fusarium oxysporum f. sp. lycopersici in tomato

Fungal pathogenic attacks are one of the major threats to the growth and productivity of crop plants. Currently, instead of synthetic fungicides, the use of plant growth-promoting bacterial endophytes has been considered intriguingly eco-friendly in nature. Here, we aimed to investigate the in vitro and in vivo antagonistic approach by using seed-borne endophytic Bacillus amyloliquefaciens RWL-1 against pathogenic Fusarium oxysporum f. sp. lycopersici. The results revealed significant suppression of pathogenic fungal growth by Bacillus amyloliquefaciens in vitro. Further to this, we inoculated tomato plants with RWL-1 and F. oxysporum f. sp. lycopersici in the root zone. The results showed that the growth attributes and biomass were significantly enhanced by endophytic-inoculation during disease incidence as compared to F. oxysporum f. sp. lycopersici infected plants. Under pathogenic infection, the RWL-1-applied plants showed increased amino acid metabolism of cell wall related (e.g., aspartic acid, glutamic acid, serine (Ser), and proline (Pro)) as compared to diseased plants. In case of endogenous phytohormones, significantly lower amount of jasmonic acid (JA) and higher amount of salicylic acid (SA) contents was recorded in RWL-1-treated diseased plants. The phytohormones regulation in disease incidences might be correlated with the ability of RWL-1 to produce organic acids (e.g., succinic acid, acetic acid, propionic acid, and citric acid) during the inoculation and infection of tomato plants. The current findings suggest that RWL-1 inoculation promoted and rescued plant growth by modulating defense hormones and regulating amino acids. This suggests that bacterial endophytes could be used for possible control of F. oxysporum f. sp. lycopersici in an eco-friendly way.

Mitochondrial Genome Analysis of Wild Rice (Oryza minuta) and Its Comparison with Other Related Species

Oryza minuta (Poaceae family) is a tetraploid wild relative of cultivated rice with a BBCC genome. O. minuta has the potential to resist against various pathogenic diseases such as bacterial blight (BB), white backed planthopper (WBPH) and brown plant hopper (BPH). Here, we sequenced and annotated the complete mitochondrial genome of O. minuta. The mtDNA genome is 515,022 bp, containing 60 protein coding genes, 31 tRNA genes and two rRNA genes. The mitochondrial genome organization and the gene content at the nucleotide level are highly similar (89%) to that of O. rufipogon. Comparison with other related species revealed that most of the genes with known function are conserved among the Poaceae members. Similarly, O. minuta mt genome shared 24 protein-coding genes, 15 tRNA genes and 1 ribosomal RNA gene with other rice species (indica and japonica). The evolutionary relationship and phylogenetic analysis revealed that O. minuta is more closely related to O. rufipogon than to any other related species. Such studies are essential to understand the evolutionary divergence among species and analyze common gene pools to combat risks in the current scenario of a changing environment.

Salvaging effect of triacontanol on plant growth, thermotolerance, macro-nutrient content, amino acid concentration and modulation of defense hormonal levels under heat stress

In this study, it was hypothesized that application of triacontanol, a ubiquitous saturated primary alcohol, at different times-before (TBHS), mid (TMHS), and after (TAHS) heat stress-will extend heat stress (HS) protection in mungbean. The effect of triacontanol on the levels of defense hormones abscisic acid (ABA) and jasmonic acid (JA) was investigated along with the plant growth promotion, nutrient and amino acid content with and without heat stress. Heat stress caused a prominent reduction in plant growth attributes, nutrient and amino acid content, which were attributed to the decreased level of ABA and JA. However, application of triacontanol, particularly in the TBHS and TMHS treatments, reversed the deleterious effects of HS by showing increased ABA and JA levels that favored the significant increase in plant growth attributes, enhanced nutrient content, and high amount of amino acid. TAHS, a short-term application of triacontanol, also significantly increased ABA and JA levels and thus revealed important information of its association with hormonal modulation. The growth-promoting effect of triacontanol was also confirmed under normal growth conditions. To the best of our knowledge, this study is the first to demonstrate the beneficial effects of triacontanol, with or without heat stress, on mungbean and its interaction with or regulation of the levels of defense hormones.

Endophytes from medicinal plants and their potential for producing indole acetic acid, improving seed germination and mitigating oxidative stress.

Medicinal plants have been used by marginal communities to treat various ailments. However, the potential of endophytes within these bio-prospective medicinal plants remains unknown. The present study elucidates the endophytic diversity of medicinal plants (Caralluma acutangula, Rhazya stricta, and Moringa peregrina) and the endophyte role in seed growth and oxidative stress. Various organs of medicinal plants yielded ten endophytes, which were identified as Phoma sp. (6 isolates), Alternaria sp. (2), Bipolaris sp. (1), and Cladosporium sp. (1) based on 18S rDNA sequencing and phylogenetic analysis. The culture filtrates (CFs; 25%, 50%, and 100% concentrations) from these endophytes were tested against the growth of normal and dwarf mutant rice lines. Endophytic CF exhibited dose-dependent growth stimulation and suppression effects. CF (100%) of Phoma sp. significantly increased rice seed germination and growth compared to controls and other endophytes. This growth-promoting effect was due to the presence of indole acetic acid in endophytic CF. The gas chromatography/mass spectrometry (GC/MS) analysis showed the highest indole acetic acid content ((54.31±0.21) μmol/L) in Bipolaris sp. In addition, the isolate of Bipolaris sp. exhibited significantly higher radical scavenging and anti-lipid peroxidation activity than the other isolates. Bipolaris sp. and Phoma sp. also exhibited significantly higher flavonoid and phenolic contents. The medicinal plants exhibited the presence of bio-prospective endophytic strains, which could be used for the improvement of crop growth and the mitigation of oxidative stresses.摘要目的探讨药用植物内生菌的多样性及其在种子生长和氧化应激中的作用。方法从三种药用植物(Caralluma acutangula、Rhazya stricta 和 Moringa peregrina)中提取内生菌;基 于18S rDNA 测序和系统发育分析鉴定分离得到 的内生菌株;以正常和矮化突变体水稻品系为对 照,比较不同浓度的内生菌培养滤液(CF)对水 稻种子的发芽和生长的影响;通过气相色谱-质谱 分析CF 中的有效活性成分。结论从药用植物中共获得10 种内生菌,包括茎点霉 属6 株、链格孢属2 株、双极霉属1 株和枝孢霉 属1 株。CF 表现出剂量依赖性的生长刺激和抑制 作用。与对照和其他内生菌相比,100%的茎点霉 菌CF 显著促进了水稻种子的发芽和生长;双极 霉中的吲哚乙酸含量最高,并表现出比其更高的 自由基清除和抗脂质过氧化活性;双极霉菌和茎 点霉菌的类黄酮和酚类成分较高。综上所述,药 用植物中存在内生菌株,其可以用于改善作物生 长和减轻氧化应激。