Genome-wide identification of <i>U-box</i> gene family and expression analysis under abiotic stresses in <i>Salvia miltiorrhiza</i>

Plant U-box (PUB) E3 ubiquitin ligases play important roles in hormone signaling pathways and in response to different abiotic stresses, but little is known about U-box genes in Danshen (root of Salvia miltiorrhiza Bunge). Here, we identified and characterized 70 SmPUB genes based on its genome sequence. Phylogenetic analysis of U-box genes from S. miltiorrhiza and Arabidopsis suggested that they can be clustered into seven subgroups (I–VII). Typical U-box domains were found in all identified SmPUB genes through the analysis of conserved motifs. Moreover, qRT-PCR was applied to analyze the relative expression levels of U-box genes in S. miltiorrhiza roots and leaves under PEG-induced water deficit and salt stresses. Results revealed that the SmPUB genes exhibited stronger response to drought than to salt stress. To the best of our knowledge, this report is the first to perform genome-wide identification and analysis of the U-box gene family in S. miltiorrhiza, and the results provide valuable information for better understanding of the function of U-box in S. miltiorrhiza.     

Identification of <i>PtGai</i> (a DELLA protein) in trifoliate orange and expression patterns in response to drought stress

Gibberellins (GAs) are an important hormone in regulating plant growth and development, and DELLA protein is an essential negative regulator of GA signal transduction. The aim of the study was to clone a GA-inhibiting protein DELLA from trifoliate orange (Poncirus trifoliata L. Raf.) and to analyze the bioinformations and expression patterns of the protein gene in tissues and in response to drought stress. A DELLA protein was isolated from trifoliate orange and named as PtGai (Genebank number: MZ170959). The PtGai protein had 1731 bp open reading frames, along with 576 amino acid codes, and also grouped with sweet orange (XM_006430552.4). The PtGai protein sequence was 65% homology with the sequences of DELLA proteins in other plant families. PtGai protein existed in the nucleus based on the prediction of subcellular localization. PtGai protein could be expressed in roots, stems, and leaves, along with the highest expression in stems. PtGai was upregulated by drought stress in leaves and roots, along with the decrease of root total GA concentration and the inhibition of shoot and root biomass production. It indicated the characteristics of PtGai protein and the roles of PtGai in GA synthesis and plant growth.     

A <i>Brachypodium distachyon</i> calcineurin B-like protein-interacting protein kinase,BdCIPK26, enhances plant adaption to drought and high salinity stress

As sessile organisms, plants possess a complex system to cope with environmental changes. Ca²⁺ functions as a vital second messenger in the stress signaling of plants, and the CBL-interacting protein kinases (CIPKs) serve as essential elements in the plant Ca²⁺ signaling pathway. In this study, calcineurin B-like protein-interacting protein kinase 26 (BdCIPK26) from Brachypodium distachyon was characterized. Overexpression of BdCIPK26 enhanced tolerance to drought and salt stress of transgenic plants. Further investigations revealed that BdCIPK26 participated in abscisic acid (ABA) signaling, conferred hypersensitivity to exogenous ABA in transgenic plants, and promoted endogenous ABA biosynthesis. Moreover, BdCIPK26 was found to maintain ROS homeostasis in plants under stress conditions. Therefore, this study indicates that BdCIPK26 functions as a positive regulator in drought and salt stress response.     

Overexpression of a sugarcane<i> ScCaM</i> gene negatively regulates salinity and drought stress responses in transgenic <i>Arabidopsis thaliana</i>

Calmodulin (CaM) proteins play a key role in signal transduction under various stresses. In the present study, the effects of a sugarcane ScCaM gene (NCBI accession number: GQ246454) on drought and salt stress tolerance in transgenic Arabidopsis thaliana and Escherichia coli cells were evaluated. The results demonstrated a significant negative role of ScCaM in the drought and salt stress tolerance of transgenic lines of A. thaliana, as indicated by the phenotypes. In addition, the expression of AtP5CS and AtRD29A, two genes tightly related to stress resistance, was significantly lower in the overexpression lines than in the wild type. The growth of E. coli BL21 cells expressing ScCaM showed weaker tolerance under mannitol and NaCl stress. Taken together, this study revealed that the ScCaM gene plays a negative regulatory role in both mannitol and NaCl stresses, and it possibly exerts protective mechanisms common in both prokaryotes and eukaryotes under stress conditions.     

<i>Agrobacterium rhizogenes</i> vs auxinic induction for <i>in vitro</i> rhizogenesis of <i>Prosopis chilensis</i> and <i>Nothofagus alpina</i>

The induction and improvement of in vitro rhizogenesis of microshoots of Prosopis chilensis (Mol.) Stuntz and Nothofagus alpina (Poep. et Endl. Oerst.) were compared using Agrobacterium rhizogenes (Ar) versus indole-3-butyric acid (IBA) in the culture media. Microshoots of P. chilensis (1-2 cm length), coming from in vitro grown seedlings, were cultivated in a modified Broadleaved Tree Medium (BTMm) containing half salt concentration of macronutrients and 0.05 mg.L^-1 benzilaminopurine (BAP). After 30 days, microshoots with 2-4 leaves were selected and cultured in BTMm-agar in presence or abscense of Ar and in combination with IBA. For N. alpina, the apical shoots with the first 2 true leaves, from 5 weeks old seedlings, were cultured in the abovementioned medium, but with 0.15 mg.L^-1 of BAP. After 2 months, microshoots with 2-3 leaves were selected and cultured in BTMm-agar, supplemented with 5 mg.L^-1 IBA or in liquid BTMm on perlite and, in the presence or absence of A. rhizogenes (Ar) and in combination with 3 mg.L^-1 IBA. Rooting in P. chilensis reached 100.0% when Ar infection was produced in the presence of IBA, increasing both, the number and dry weight of roots. In N. alpina, 90.0% of rooting efficiency was obtained when Ar infection was produced in liquid culture and in the absence of auxin.     

Salinity induced anatomical and morphological changes in <i>Chloris gayana</i> Kunth roots

Chloris gayana Kunth is a grass species valuable as forage which was introduced into Argentina to be used as pasture in saline soils of subtropical and warm-temperate zones, given its good adaptability to drought, salinity and mild freezing. However, its tolerance varies according to the cultivar. In tetraploid cultivars, important reductions in yield have been observed. Here, a study of the variations produced on the root and stem system by salinity at different NaCl concentrations (0, 150 y 250 mM) was performed in the Boma cultivar, with the aim of determining the anatomical and morphological alterations produced by the salt excess. Plants cultivated with the highest level of salinity showed, in the whole, significant differences in the measured variables. A diminution in absolute values of the variables and a major reduction in vascular tissue dimensions were observed, which suggests that the lack of tolerance to salt stress could be related to a deficient adaptation to absorb and transport water and nutrients from the roots.     

Role of foliar spray of plant growth regulators in improving photosynthetic pigments and metabolites in <i>Plantago ovata</i> (Psyllium) under salt stress–A field appraisal

Salinity is one of the major abiotic factors that limit the growth and productivity of plants. Foliar application of plant growth regulators (PGRs) may help plants ameliorate the negative impacts of salinity. Thus, a field experiment was conducted at the Botanical Garden University of Balochistan, Quetta, to explore the potential role of PGRs, i.e., moringa leaf extract (MLE; 10%), proline (PRO; 1 µM), salicylic acid (SA; 250 µM), and thiourea (TU; 10 mM) in ameliorating the impacts of salinity (120 mM) on Plantago ovata, an important medicinal plant. Salinity hampered plant photosynthetic pigments and metabolites but elevated oxidative parameters. However, foliar application of PGRs enhanced photosynthetic pigments, including Chl b (21.11%), carotenoids (57.87%) except Chl a, activated the defense mechanisms by restoring and enhancing the metabolites, i.e., soluble sugars (49.68%), soluble phenolics (33.34%), and proline (31.47%), significantly under salinity stress. Furthermore, foliar supplementation of PGRs under salt stress led to a decrease of about 43.02% and 43.27% in hydrogen peroxide and malondialdehyde content, respectively. Thus, PGRs can be recommended for improved photosynthetic efficiency and metabolite content that can help to get better yield under salt stress, with the best and most effective treatments being those of PRO and MLE to predominately ameliorate the harsh impacts of salinity.     

Response of <i>MaHMA2</i> gene expression and stress tolerance to zinc stress in mulberry (<i>Morus alba</i> L.)

HMA2 (heavy metal ATPase 2) plays a crucial role in extracellular and intracellular Zn²⁺ transport across biomembranes, maintaining ion homeostasis, and playing an important role in the normal physiological metabolism, growth, and development of plants. In our study, a novel HMA2 gene, named MaHMA2, was isolated and cloned from white mulberry (Morus alba L.). The gene sequence obtained was 1,342 bp long, with an open reading frame of 1,194 bp, encoding a protein of 397 amino acids, with a predicted molecular mass of 42.852 kD and an isoelectric point of 7.53. This protein belonged to the PIB-type ATPase transport protein family. We analyzed the expression of the MaHMA2 gene by quantitative real-time PCR. The results showed that the level of MaHMA2 gene expression decreased to a Zn concentration of 800 mg/kg. Malondialdehyde and proline levels increased and responded to increasing Zn when the MaHMA2 gene was silenced, whereas the activities of peroxidase and superoxide dismutase tended to increase in response to increasing Zn²⁺ ion stress concentrations but were lower in the gene-silenced plants. These findings suggested that the MaHMA2 gene played an active role in the tolerance response of mulberry to Zn stress.     

Association between preterm birth risk and polymorphism and expression of the DNA repair genes <i>OGG1</i> and <i>APE1</i> in Saudi women

Genomic instability and mutations caused by increases in oxidative stress during pregnancy can damage the fetoplacental unit and can upshot preterm birth. Oxidative damage to DNA may possibly be involved in etiology of preterm birth (PTB) which can be repaired by DNA repair gene. In the present study, we assessed the association of base excision repair gene family by analyzing the association of single nucleotide polymorphisms and genes expression in 8-oxoguanine glycosylase-1 (OGG1) and apurinic-apyrimidinic endonuclease 1 (APE1) genes with risk of preterm birth in Saudi women. We analyzed genotypes of four single nucleotide polymorphisms (SNPs) (rs1052133, rs293795, rs2072668 and rs2075747) in OGG1 gene and three SNPs (rs1130409, rs3136814, and rs3136817) in APE1 gene using TaqMan Genotyping assay kits in 50 pairs of preterm cases and individually matched controls. Also, gene expression level was explored by RT-PCR in 10 pairs of preterm placental tissues and individually matched normal placental tissues. Two OGG1 SNP, rs1052133 (OR=0.497; c2=1.11; p=0.292) and rs2072668 (OR=0.408; c2=1.90; p=0.167) and one APE1 SNP rs3136817 (OR=0.458; c2=0.40; p=0.527) showed nonsignificant protective effect against PTB development. The expression of both genes under study was found lower in the PTB patients. Genotype and allele frequencies of both gene SNPs did not show any association with the risk of preterm delivery in Saudi women (P˃0.05). However, synthesis and release of OGG1 and APE1 proteins decreased in preterm placental tissues compared to term delivery reflects the probability of being one of the mechanisms leading to preterm birth.     

Screening and evaluation of chilli (<i>Capsicum annuum</i> L.) genotypes for waterlogging tolerance at seedling stage

Waterlogging is an illustrious abiotic stress and the constrictions it enforces on plant roots have negative effects on growth and development. This study was undertaken to investigate waterlogging stress tolerant potential in chilli (Capsicum annum L.) genotypes through evaluating morphological, physiological, biochemical and anatomical parameters. Thirty-five days old seedlings of 10 chilli genotypes were exposed to waterlogging stress maintaining water height 3–5 cm over the soil surface artificially for three days. This duration (36–38 DAE) was termed as waterlogging period, and subsequent withdrawal of waterlogging condition (39–45 DAE) was regarded as a recovery phase. Based on their survival performance, two tolerant genotypes viz., SRC-517 and BARI morich-2 and two susceptible genotypes viz., AHM-206 and RI-1(6) were selected for studying stress tolerance mechanism. Under waterlogging, however, both genotypes (tolerant and susceptible) exhibited reduced root shoot length, dry weight ratio, petiole weight and leaf area, and noticeable reduction regarding these parameters was observed in susceptible genotypes. Moreover, tolerant genotypes displayed a higher recovery than susceptible genotypes after removal of waterlogging stress. Lower reduction of leaf area and photosynthetic pigments as well as higher reduction of relative water content (RWC) were noticed in susceptible genotypes. Higher accumulation of proline and total antioxidant capacity (TAC) during waterlogging condition in tolerant genotypes suggested lower oxidative damage. Although both genotypes lost total soluble sugar (TSS) relative to control at waterlogging stress, better performance was recorded in tolerant genotypes. During the period after the removal of extra water, a similar genotypic response in terms of TSS gain was seen. Undoubtedly, under flooding conditions, the development of aerenchyma cells in tolerant genotypes is a means of tolerance mechanism for long-term survival. Thus, the morpho-physiological and biochemical changes help to understand the tolerance mechanism in chilli under waterlogging stress.     

Expression analysis of <i>OsSERK</i>, <i>OsLEC1</i> and <i>OsWOX4</i> genes in rice (<i>Oryza</i> sativa L.) callus during somatic embryo development

Somatic embryogenesis is an asexual reproduction process that occurs in many plant species, including rice. This process contains several totipotency markers such as Somatic Embryogenesis Receptor-like Kinase (SERK), Leafy Cotyledon1 (LEC1) and WUSCHEL-Related Homeobox4 (WOX4) and also a helpful model for embryo development and clones and transformations. Here, we report the gene expression during somatic embryo development correlates with regeneration frequency in 14 Javanica rice (pigmented and non-pigmented) using modifified N6 media supplemented with Kinetin (2.0 mg/L) and NAA (1.0 mg/L). Although there have been advances in understanding the genetic basis of somatic embryogenesis in other varieties, rice is still unexplored, especially during somatic embryo development. Moreover, for the formation of callus induction from immature embryos, 2,4-D (2.0 mg/L, 3.0 mg/L) was used. This study analysed the gene expression of OsSERK, OsWOX4 and OsLEC1 genes through RT-PCR analysis. Higher expression of the OsLEC1 gene indicates that their function may correlate in the in vitro with the high response of rice after transfer to regeneration media. This study found that rice varieties of pigmented rice (MS Pendek and Gogoniti II) and non-pigmented rice (Pandan Ungu) showed high regeneration frequency, showing higher OsLEC1 expression than other varieties because OsLEC1 promotes the maturation of somatic embryos in plant regeneration on day 14. However, the contrast with Genjah nganjuk may be effective because of other regulatory genes. RT-PCR analysis showed OsSERK had less expression level than OsLEC1 and OsWOX4 in the varieties, which correlate with the percentage of plant regeneration, but not for Gogoniti II. In conclusion, the higher percentage of plant regeneration correlates with the higher expression level of OsLEC1 at day 14 of media regeneration of rice.     

Isolation and molecular characterization of a <i>cax</i> gene from <i>Capsella bursa-pastoris</i>

A new cation exchangers (CAXs) gene was cloned and characterized from Capsella bursapastoris by rapid amplification of cDNA ends (RACE). The full-length cDNA sequence of cax from C. bursa-pastoris (designated as Cbcax51) was 1754 bp containing a 1398 bp open reading frame encoding a polypeptide of 466 amino-acid residues with a calculated molecular mass of 50.5 kDa and an isoelectric point of 5.69. The predicted CbCAX51 contained an IMP dehydrogenase/GMP reductase domain, two Na⁺/Ca²⁺ exchanger protein domains. Comparative and bioinformatics analyses revealed that CbCAX51 showed extensive homology with CAX from other plant species. The expression analysis by different treatments indicated that Cbcax51 could be activated by cold triggering and was related to the cold acclimation process, but its expression is regulated negatively by drought and not affected by ABA or salt.