Plant growth-promoting phyllosphere bacteria mediated ACC oxidase gene regulation in groundnut to mitigate drought stress

We investigated the genetics of ethylene biosynthesis and its linkage to the RFLPs of the ACC oxidase and synthase genes in melon ( Cucumis melo L.). The results suggested that the A(0) and B(0) fragments of RFLP-MEL1 of the ACC oxidase gene were two alleles from a single locus, as were the B and C fragments of RFLP-MEACS1 of the ACC synthase gene. The B(0) allele seemed to be partially dominant over the A(0) allele, whereas B and C alleles appeared to map to quantitative trait loci (QTLs), which most likely contributed to ethylene production. Both RFLPs were linked to ethylene production rates, but they were not linked to each other. The interaction effects of the ACC oxidase and synthase genes on ethylene production were revealed by segregation of RFLP-MEL1 and RFLP-MEACS1. The results of single-copy-reconstruction assays suggested that the ACC oxidase gene is a single copy, whereas the ACC synthase gene is a component of a multigene family in the melon genome. The abscission phenotype appeared to be controlled by an independent locus, with the abscission (full-slip) allele dominant over the non-abscission (not full-slip) allele. These results may facilitate efforts toward mapping the quantitative trait loci (QTLs) of ethylene production. The RFLPs may be used in marker-assisted selection in developing melons with a more-desirable low ethylene production rate for enhancing postharvest storage life.

ACC oxidase (ACO) genes in Trifolium occidentale (L.) and their relationship to ACO genes in white clover (T. repens L.) and T. pallescens (L.)

Proline accumulation and glutathione reductase activity induced by drought-tolerant rhizobacteria as potential mechanisms to alleviate drought stress in Guinea grass

Does the enhanced tolerance of arbuscular mycorrhizal plants to water deficit involve modulation of drought‐induced plant genes?

Chapter 22 - Bacterial alleviation of drought stress in plants: recent advances and future challenges

Exogenous spermidine regulates the anaerobic enzyme system through hormone concentrations and related-gene expression in Phyllostachys praecox roots under flooding stress

Differential regulation of two 1-aminocyclopropane-1-carboxylate oxidase (ACO) genes, including the additionally cloned DcACO2, during senescence in carnation flowers

The effects of drought on growth and seed quality of oilseed crops are of crucial importance in edible oil production due to its pivotal role in sustainable agriculture. Plant growth-promoting bacteria (PGPB) can improve crop yield by promoting plant growth under various environmental conditions. In the present study, the physiological responses, growth, and seed quality of three camelina doubled haploid lines (DH51, DH69, and DH104) were assessed upon their exposure to two irrigation regimes at the presence of Micrococcus yunnanensis during their reproductive phase. The results showed that the investigated parameters of camelina were affected by genotype, irrigation regimes, and PGPB. Drought decreased crop yield as measured by silique length, silique, and seed number and 1000-weight seed. PGPB significantly decreased the adverse effects of stress consistent by increasing the branches per plant and root length. Drought also caused a significant enhancement in the hydrogen peroxide and malondialdehyde contents, but the PGPB-inoculated plants showed lower contents of both compounds. Relative water content significantly reduced in plant grown under stress but inoculation enhanced the potential of water retaining in plants under stress and non-stress conditions. Drought stress and PGPB elevated proline and total soluble carbohydrate content in genotypes. Drought stress had no significant effect on photosynthetic pigments content of genotypes while inoculation apparently moderated negative impact of drought with enhancement of pigments content. The obtained results were responsible for metabolic changes occurring in response to stress. PGPB improved the plant drought-tolerance by enhancing its physiological traits. The fatty acid profile showed some variations among camelina genotypes under drought stress and PGPB inoculation. Upon symbiosis association, an increase was observed in major constituents of polyunsaturated acids, linoleic and linolenic acids, and a significant increase in oleic acid as a main monounsaturated acid. They also altered another major constituent, gadeolic acid, under water deficit stress and/ or with PGPB. Both drought stress and PGPB decreased the poly unsaturated fatty acids/mono unsaturated fatty acids ratio. In general, there was a significant difference among camelina lines in terms of seed yield and quality in response to drought. Also, it strongly suggested that PGPB application can be a positive strategy to mitigate drought stress and increase crop yield.

Caterpillar-induced ethylene emissions play an important role in plant-herbivore interactions. The ethylene burst that ensues after attack exceeds wound-induced ethylene emissions, but the mechanisms responsible remain unknown. Adding larval oral secretions (OS) to wounds mimics this ethylene burst. We demonstrate that fatty acid-amino acid conjugates are the responsible elicitors in Manduca sexta OS, and identify genes that are important in OS-elicited ethylene biosynthesis and perception in the larvae's host, Nicotiana attenuata, by examining the consequences of gene silencing on OS-elicited ethylene emissions, as quantified by photo-acoustic spectroscopy. OS elicitation increased transcript accumulation of ACC synthase (ACS), virus-induced gene silencing of ACS halved the OS-elicited ethylene release, and ACC supplementation to ACS-silenced plants restored ethylene emissions, demonstrating that ACS activity limits the rate of release. Silencing three wound- or OS-elicited ACC oxidase (ACO) genes with an ACO consensus fragment abolished the OS-elicited ethylene release. Virus-induced gene silencing of each ACO individually revealed that only NaACO2a and NaACO3 regulate the OS-elicited ethylene release. Transforming plants with various etr1-1 constructs rendered them differentially 'deaf' to ethylene, and dramatically increased the OS-elicited ethylene burst, largely without regulating the transcripts of biosynthetic genes. The volume of the OS-elicited ethylene 'scream' was proportional to the plant's deafness, as determined by 1-MCP treatments. We conclude that the OS-elicited ethylene burst is tuned by a tag-team of transcriptional responses and ethylene perception. Ethylene signaling is shown to be essential in regulating two traits that are important in the N. attenuata-M. sexta interaction: OS-induced nicotine levels and floral longevity.

Drought is one of the most detrimental factors that causes significant effects on crop development and yield. However, the negative effects of drought stress may be alleviated with the aid of exogenous melatonin (MET) and the use of plant-growth-promoting bacteria (PGPB). The present investigation aimed to validate the effects of co-inoculation of MET and Lysinibacillus fusiformis on hormonal, antioxidant, and physio-molecular regulation in soybean plants to reduce the effects of drought stress. Therefore, ten randomly selected isolates were subjected to various plant-growth-promoting rhizobacteria (PGPR) traits and a polyethylene-glycol (PEG)-resistance test. Among these, PLT16 tested positive for the production of exopolysaccharide (EPS), siderophore, and indole-3-acetic acid (IAA), along with higher PEG tolerance, in vitro IAA, and organic-acid production. Therefore, PLT16 was further used in combination with MET to visualize the role in drought-stress mitigation in soybean plant. Furthermore, drought stress significantly damages photosynthesis, enhances ROS production, and reduces water stats, hormonal signaling and antioxidant enzymes, and plant growth and development. However, the co-application of MET and PLT16 enhanced plant growth and development and improved photosynthesis pigments (chlorophyll a and b and carotenoids) under both normal conditions and drought stress. This may be because hydrogen-peroxide (H2O2), superoxide-anion (O2-), and malondialdehyde (MDA) levels were reduced and antioxidant activities were enhanced to maintain redox homeostasis and reduce the abscisic-acid (ABA) level and its biosynthesis gene NCED3 while improving the synthesis of jasmonic acid (JA) and salicylic acid (SA) to mitigate drought stress and balance the stomata activity to maintain the relative water states. This may be possible due to a significant increase in endo-melatonin content, regulation of organic acids, and enhancement of nutrient uptake (calcium, potassium, and magnesium) by co-inoculated PLT16 and MET under normal conditions and drought stress. In addition, co-inoculated PLT16 and MET modulated the relative expression of DREB2 and TFs bZIP while enhancing the expression level of ERD1 under drought stress. In conclusion, the current study found that the combined application of melatonin and Lysinibacillus fusiformis inoculation increased plant growth and could be used to regulate plant function during drought stress as an eco-friendly and low-cost approach.

An ACC Oxidase Gene Essential for Cucumber Carpel Development

Ethylene is important during the berry development and in the last stages of rachis development or rachis senescence. Since grapes develop in a cluster that comprises both the fruit berry and the nonfruit rachis, we measured the release of ethylene from both tissues. Detached berries from Vitis vinifera ‘Ruby Seedless’ and ‘Thompson Seedless’ showed that ethylene release peaks at the beginning of berry development and at veraison. Ethylene production in the rachis was higher than that in the berry and had an obvious peak before harvest in ‘Thompson Seedless’. In both cultivars, ethephon treatment induced ethylene production in the rachis but not in the berry. Expression of 1-aminocyclopropane-1-carboxylate (ACC) synthase (ACS) and ACC oxidase (ACO) genes showed diverse temporal and spatial patterns in ‘Thompson Seedless’ and ‘Ruby Seedless’. For most gene family members, the low ACS expression levels were observed in berry and rachis. Expression levels of most of the ACS and ACO genes did not correlate with ethylene released in the same organ. The transcriptional level of VvACS1 did correlate with ethylene evolution in rachis of ‘Thompson Seedless’ during berry development and storage, which suggested that VvACS1 may have important roles in rachis senescence. In berries of ‘Thompson Seedless’ and ‘Ruby Seedless’, the transcriptional levels of VvACO1, VvACS2, and VvACS6 coincided with ethylene production, indicating possible roles in berry development. Expression of VvACS2–VvACO9 and VvACO1–VvACO3 was not consistent with ethylene production during storage or in response to ethephon treatment, which suggests that the expression of ACS and ACO was affected by other stress factors after harvest.

In an effort towards understanding the biochemical properties and physiological functions of 1-aminocyclopropane-1-carboxylic acid (ACC) oxidase homologues, we have isolated three ACC oxidase clones from sunflower (Helianthus annuus) seedlings. ACCO1 is a cDNA clone while ACCO2 and ACCO3 and reverse transcriptase-polymerase chain reaction clones. Southern analysis indicated the existence of at least three members in the sunflower ACC oxidase gene family. Expression studies showed that ACCO3 was equally expressed in leaves, hypocotyl, and roots of sunflower seedlings, but it constituted only a small amount of the total ACC oxidase transcripts. In contrast, ACCO1 and ACCO2 were differentially expressed in these organs. ACCO1 mRNA was most abundant in roots, whereas ACCO2 was the major homologue in leaves and in hypocotyl. The levels of total ACC oxidase transcripts in these organs were also determined. High ACC oxidase transcript levels were associated with tissue containing rapidly dividing cells. Wounding and silver ion treatments of hypocotyls increased ACC oxidase mRNA levels and ACC oxidase activity; these events being consistent with the increases in ethylene production. In contrast, ACC oxidase protein levels were not affected by these treatments, suggesting that either a translational regulation and/or a rapid turn-over of the protein is involved in both wound- and silver ion-induced gene expression. Contrary to data in the literature, we found that auxins, ethephon and ACC did not affect ACC oxidase mRNA levels in sunflower hypocotyls. The complexity of ACC oxidase regulation and the significance of organ differential expression of ACC oxidase genes are discussed.

ACC (1-aminocyclopropane-1-carboxylate) oxidase genes are differentially expressed in melon during development and in response to various stresses. We investigated the molecular basis of their transcription by analyzing the 5' untranslated regions of the ACC oxidase genes CM-ACO1 and CM-ACO3. In order to determine how their temporal and spatial expression patterns were established, we fused the promoter regions of CM-ACO1 (726 bp) and CM-ACO3 (2260 bp) to the beta-glucuronidase (GUS) reporter gene and examined their regulation in transgenic tobacco plants. The CM-ACO1 promoter was able to drive GUS expression in response to wounding, and to treatment with ethylene or copper sulfate. It was also rapidly induced (8-12 h postinoculation) in tobacco leaves inoculated with the hypersensitive response (HR)-inducing bacterium Ralstonia solanacearum. Expression was also observed during compatible interactions but was delayed. In contrast, the CM-ACO3 promoter was not expressed in response to infection, but was up-regulated during flower development. Both promoters were regulated during leaf senescence but in different patterns. The CM-ACO1-driven GUS activity increased sharply concomitantly with the onset of chlorophyll breakdown, while the CM-ACO3 promoter drove strong GUS expression in green, fully expanded leaves and this declined at the onset of senescence. This result is consistent with the expression patterns of these two genes in senescent melon leaves. These data suggest that the regulation of expression of CM-ACO1 is related preferentially to stress responses, whereas CM-ACO3 seems to be associated with developmental processes. The possible role of ethylene is discussed, particularly in the regulation of the CM-ACO1 gene in response to stress and during senescence.

Plant growth promoting bacteria (PGPB) are associated with plant roots and augment plant productivity and immunity; however, recent work by several groups has shown that PGPB also elicit the so-called “induced systemic tolerance” to drought stress. This work aimed to evaluate the effect of Gluconacetobacter diazotrophicus Pal5 inoculation on the mitigation of drought stress in red rice (Oryza sativa L.). The experiment treatments of the red rice cultivar inoculated and uninoculated with G. diazotrophicus, and cultivated with and without water restriction. Physiological, biochemical, and molecular analyses of plant tissues were carried out, along with measurements of biomass and seed yield components. The plants showed a positive response to inoculation with G. diazotrophicus, with promotion of growth and induction of tolerance to drought. Increase in plant biomass and higher levels of gas exchange and osmoprotectant solutes were observed in shoots. The bacterial inoculation increased the yield and positively regulated some defense genes against the water deficit in plants. G. diazotrophicus Pal5 strain inoculation was beneficial for red rice plants, suppressing the various deleterious effects of drought stress and increasing the biomass and seed yield components.