Regulation and physiological function of proteins for heat tolerance in cowpea (Vigna unguiculata) genotypes under controlled and field conditions.

The expression of heat shock proteins is considered a central adaptive mechanism to heat stress. This study investigated the expression of heat shock proteins (HSPs) and other stress-protective proteins against heat stress in cowpea genotypes under field (IT-96D-610 and IT-16) and controlled (IT-96D-610) conditions. Heat stress response analysis of proteins at 72 h in the controlled environment showed 270 differentially regulated proteins identified using label-free quantitative proteomics in IT-96D-610 plants. These plants expressed HSPs and chaperones [BAG family molecular chaperone 6 (BAG6), Multiprotein bridging factor1c (MBF1C) and cold shock domain protein 1 (CSDP1) in the controlled environment]. However, IT-96D-610 plants expressed a wider variety of small HSPs and more HSPs in the field. IT-96D-610 plants also responded to heat stress by exclusively expressing chaperones [DnaJ chaperones, universal stress protein and heat shock binding protein (HSBP)] and non-HSP proteins (Deg1, EGY3, ROS protective proteins, temperature-induced lipocalin and succinic dehydrogenase). Photosynthesis recovery and induction of proteins related to photosynthesis were better in IT-96D-610 because of the concurrent induction of heat stress response proteins for chaperone functions, protein degradation for repair and ROS scavenging proteins and PSII operating efficiency (Fq'/Fm') than IT-16. This study contributes to identification of thermotolerance mechanisms in cowpea that can be useful in knowledge-based crop improvement.

Proteome changes and associated physiological roles in chickpea (Cicer arietinum) tolerance to heat stress under field conditions.

Interrogative proteome analyses are used to identify and quantify the expression of proteins involved in heat tolerance and to identify associated physiological processes in heat-stressed plants. The objectives of the study were to identify and quantify the expression of proteins involved in heat tolerance and to identify associated physiological processes in chickpea (Cicer arietinum L.) heat-tolerant (Acc#7) and sensitive genotype (Acc#8) from a field study. Proteomic and gene ontological analyses showed an upregulation in proteins related to protein synthesis, intracellular traffic, defence and transport in the heat-tolerant genotype compared to the susceptible one at the warmer site. Results from KEGG analyses indicate the involvement of probable sucrose-phosphate synthase (EC 2.4.1.14) and sucrose-phosphate phosphatase (EC 3.1.3.24) proteins, that were upregulated in the heat-tolerant genotype at the warmer site, in the starch and sucrose pathway. The presence of these differentially regulated proteins including HSP70, ribulose bisphosphate carboxylase/oxygenase activase, plastocyanin and protoporphyrinogen oxidase suggests their potential role in heat tolerance, at flowering growth stage, in field-grown chickpea. This observation supports unaltered physiological and biochemical performance of the heat-tolerant genotypes (Acc#7) relative to the susceptible genotype (Acc#8) in related studies (Makonya et al. 2019). Characterisation of the candidate proteins identified in the current study as well as their specific roles in the tolerance to heat stress in chickpea are integral to further crop improvement initiatives.

Melatonin Regulatory Mechanisms and Phylogenetic Analyses of Melatonin Biosynthesis Related Genes Extracted from Peanut under Salinity Stress.

Melatonin improves the tolerance of plants to various environmental stresses by protecting plant cells against oxidative stress damage. The objective of the current study was to determine whether exogenous melatonin (MT) treatments could help protecting peanut (Arachis hypogaea) seedlings against salinity stress. This was achieved by investigating enzymatic and non-enzymatic antioxidant systems and the expression of melatonin biosynthesis related genes in response to salinity stress with or without exogenous MT. The results showed a significant increase in the concentrations of reactive oxygen species (ROS) in peanut seedlings under salinity stress. The exogenous application of melatonin decreased the levels of ROS through the activation of antioxidant enzymes in peanut seedlings under salinity stress. Transcription levels of melatonin biosynthesis related genes such as N-acetylserotonin methyltransferase (ASMT1, ASMT2, ASMT3), tryptophan decarboxylase (TDC), and tryptamine 5-hydroxylase (T5H) were up-regulated with a 150 µM melatonin treatment under salinity stress. The results indicated that melatonin regulated the redox homeostasis by its ability to induce either enzymatic or non-enzymatic antioxidant systems. In addition, phylogenetic analysis of melatonin biosynthesis genes (ASMT1, ASMT2, ASMT3, TDC, T5H) were performed on a total of 56 sequences belonging to various plant species including five new sequences extracted from Arachis hypogaea (A. hypogaea). This was based on pairwise comparison among aligned nucleotides and predicted amino acids as well as on substitution rates, and phylogenetic inference. The analyzed sequences were heterogeneous and the A. hypogaea accessions were primarily closest to those of Manihot esculenta, but this needs further clarification.

The Impact of Drought Stress on Antioxidant Responses and Accumulation of Flavonolignans in Milk Thistle (Silybum marianum (L.) Gaertn).

Biosynthesis and accumulation of flavonolignans in plants are influenced by different environmental conditions. Biosynthesis and accumulation of silymarin in milk thistle (Silybum marianum L.) were studied under drought stress with respect to the antioxidant defense system at the physiological and gene expression level. The results revealed a reduction in leaf chlorophyll, ascorbic acid, and glutathione contents. In contrast, H2O2, proline, and antioxidative enzyme activities, such as superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), ascorbate peroxidase (APX), and glutathione reductase (GR), were increased. These results confirmed that milk thistle undergoes oxidative stress under drought stress. Furthermore, transcription levels of APX, SOD, CAT, 1-Cys-Prx, and PrxQ were significantly increased in milk thistle under drought stress. Overall this suggests that protection against reactive oxygen species and peroxidation reactions in milk thistle are provided by enzymatic and non-enzymatic antioxidants. Flavonolignans from milk thistle seeds after different drought treatments were quantified by high-performance liquid chromatography (HPLC) and showed that severe drought stress enhanced the accumulation of silymarin and its components compared with seeds from the control (100% water capacity). Silybin is the major silymarin component and the most bioactive ingredient of the milk thistle extract. Silybin accumulation was the highest among all silymarin components in seeds obtained from drought-stressed plants. The expression of the chalcone synthase (CHS) genes (CHS1, CHS2, and CHS3), which are associated with the silybin biosynthetic pathway, was also increased during drought stress. These results indicated that milk thistle exhibits tolerance to drought stress and that seed derived from severe drought-stressed plants had higher levels of silymarin.

Enhancing antioxidant systems by exogenous spermine and spermidine in wheat (Triticum aestivum) seedlings exposed to salt stress.

Plants have evolved complex mechanisms to mitigate osmotic and ionic stress caused by high salinity. The effect of exogenous spermine (Spm) and spermidine (Spd) on defence responses of wheat seedlings under NaCl stress was investigated by measuring antioxidant enzyme activities and the transcript expression of corresponding genes. Exogenous Spm and Spd decreased the level of malondialdehyde, increased chlorophyll and proline contents, and modulated PSII activity in wheat seedlings under salt stress. Spermidine alleviated negative effects on CO2 assimilation induced by salt stress in addition to significantly increasing the activity and content of ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco). It appears Spd conferred salinity tolerance in wheat seedlings by enhancing photosynthetic capacity through regulation of gene expression and the activity of key CO2 assimilation enzymes. Exogenous Spm regulated activities of different antioxidant enzymes (catalase, glutathione reductase, dehydroascorbate reductase, ascorbate peroxidase, and superoxide dismutase) and efficiently modulate their transcription levels in wheat seedlings under salt stress. It is likely that Spm plays a key role in alleviating oxidative damage of salt stress by adjusting antioxidant enzyme activities in plants. In addition, exogenous Spd increased transcript level of spermine synthase under salt stress. Salinity stress also caused an increase in transcript levels of diamine oxidase (DAO) and polyamine oxidase (PAO). Exogenous Spd application resulted in a marked increase in free Spd and Spm contents under saline conditions. These results show that exogenous Spd and Spm effectively upregulated transcriptional levels of antioxidant enzyme genes and improved the defence response of plants under salt stress.