Plant Molecular Biology最新文献

Triacylglycerol (TAG) is a major component of plant-neutral lipids. Diacylglycerol acyltransferase 2 (DGAT2) plays an important role in plant oil accumulation by catalyzing the final step of the Kennedy pathway. In this study, ten DGAT2 sequences were originating from different oil crops into the TAG-deficient yeast strain H1246, to compare their enzyme activity of oil synthesis and filter out potential amino acid residue sites for directed evolution. Based on the synthesis efficiency of total lipids, TAGs, and the topology models of these DGAT2s, five possible amino acid sites were identified that may affect the synthesis of total lipids and TAGs. In the H1246 yeast expression system, HaDGAT2 significantly increased the total oil and TAG content; however, ClDGAT2 was weak in synthesizing both oil and TAG. Thus, building on HaDGAT2 and ClDGAT2, these amino acid substitutions were created by point-to-point mutating and substantially affected the oil or TAG synthesis ability of DGAT2s. Among the five amino acid substitutions, mutations at residue (3) successfully make HaDGAT2 less capable of synthesizing lipids and TAG, and ClDGAT2 more capable of synthesizing total lipids and TAG. Except mutations at residue (2), all residue mutations contributed to a weaker ability of fatty acid synthesis. In addition, ten mutant DGAT2s and two parental DGAT2s were overexpressed in tobacco leaves to reveal their lipid synthesis function. This approach helped us to authenticate the significance of these loci. In varying degrees, those mutations enhanced the ability of ClDGAT2 to synthesize lipids, attenuated the ability of HaDGAT2 to synthesize lipids, and altered preference for fatty acids in tobacco.

OsMAIL1 encodes for a rice protein of the Plant Mobile Domain (PMD) family and is strongly upregulated during floral induction in response to the presence of the florigens Heading date 3a (Hd3a) and RICE FLOWERING LOCUS T1 (RFT1). Although OsMAIL1 expression depends on the florigens, osmail1 null mutants do not show delay in flowering time, rather OsMAIL1 participates in ensuring successful reproduction. Indeed, when day temperatures reach 35 °C (7 °C higher than standard greenhouse conditions), osmail1 mutants show increased sterility due to abnormal pistil development with about half of the plants developing three styles topped by stigmas. OsMAIL1 expression correlates with that of carpel identity genes and RNA-seq of osmail1-1 mutant compared to the wt during inflorescence development showed that OsMAIL1 is required to activate carpel identity genes expression when floral meristems are about to be initiated. OsMAIL1 is a newly characterized rice gene that specifically controls carpel development under heat stress, ensuring plant female fertility in these conditions.

Soil salinity poses a significant environmental challenge for the growth and development of blueberries. However, the specific mechanisms by which blueberries respond to salt stress are still not fully understood. Here, we employed a comprehensive approach integrating physiological, metabolomic, and transcriptomic analyses to identify key metabolic pathways in blueberries under salt stress. Our findings indicate that blueberries primarily adapt to salt stress by modulating pathways associated with carbohydrate metabolism, organic acid metabolism, amino acid metabolism, and various organic compounds. Key metabolites involved in this response include sucrose, propionic acid, and palmitic acid. A total of 241 transcription factors were differentially expressed, with significant involvement from families such as AP2, Dof, GATA, WRKY, and TCP. Notably, the galactose metabolism pathway was associated with 5 DAMs and 24 DEGs, while the starch and sucrose metabolism pathway contained 5 DAMs and 23 DEGs, highlighting their crucial roles in mitigating salt stress. Overexpression of VcGolS3 in transgenic Arabidopsis conferred tolerance to salt and drought stresses, primarily evidenced by a significant increase in GolS enzyme activity and reduced ROS accumulation. This study provides valuable insights into the molecular mechanisms underlying the blueberry response to salt stress and lays the groundwork for breeding salt- and drought-tolerant blueberry varieties.

Deficiency or excess of mineral elements in the environment is a primary factor limiting crop yields and nutritional quality. Lotus (Nelumbo nucifera) is an important aquatic crop in Asia, but the mechanism for accumulating mineral nutrients and coping with nutrient deficiency/excess is still largely unknown. Here, we identified NnMTP10, a member of the cation diffusion facilitator family, by screening the cDNA library of lotus. Subcellular localization to the plasma membrane, increased manganese (Mn) and iron (Fe) tolerance and reduced metal accumulation in yeast transformants demonstrated that the protein functions as an exporter to mediate the efflux of Mn and Fe. Arabidopsis overexpressing NnMTP10 exhibited less Mn accumulation in roots, increased long-distance transport to shoots, and increased tolerance to Mn stress under high-Mn conditions. However, the accumulation and tolerance of Fe in Arabidopsis transformants are opposite to those of Mn. Further analysis revealed that excessive Fe in the root apoplast exported by NnMTP10 was sequestrated by the cell wall, thereby reducing the transport of Fe to the shoot. Correspondingly, the expression of NnMTP10 in the roots of lotus was increased under the high-Mn treatment but decreased under the high-Fe treatment. These results suggest that NnMTP10 is involved in the long-distance transport of Mn and Fe in lotus and may play a role in coordinating the adaptation to stresses caused by excessive Mn and Fe.

Z. armatum is an economically valued crop known for its rich aroma and medicinal properties. This study identified 45 members of the SQUAMOSA-PROMOTER BINDING PROTEIN LIKE (SPL) gene family in the genome of Z. armatum. Phylogenetic and collinearity analyzes demonstrated a close relationship between ZaSPLs and ZbSPLs from B subgenomes of Zanthoxylum bungeanum. Our miRNA sequencing revealed a high degree of conservation of miR156a within Z. armatum, with the za-miR156a sequence identical to miR156-5p in Arabidopsis thaliana and Citrus sinensis. Of the 45 genes identified by ZaSPLs, 21 were targeted by za-miR156a, transient co-expression experiments in N. benthamiana demonstrated the targeting relationship between za-miR156 and ZaSPL21. Furthermore, RNA-seq and qRT-PCR analysis revealed that ZaSPL genes exhibited elevated expression levels in juvenile tissues of Z. armatum. The expression of nine representative ZaSPL genes were upregulated under polyethylene glycol (PEG) and abscisic acid (ABA). Overexpression of ZaSPL21 delayed the germination of transgenic tobacco and facilitated the flowering process in transgenic N. benthamiana. Significant up-regulation in the expression levels of flowering-related genes such as NbFT1, NbPIP2;1, NbTCP1, NbCOL1, NbGI2, NbGAI1, NbCKX2, and NbARR4 was observed in transgenic plants, suggesting that ZaSPL21 may stimulate plant flowering by regulation of these genes. Furthermore, ZaSPL21 also increased the germination speed of transgenic tobacco seeds during drought and salt stress conditions, and improved the salt tolerance of transgenic seedlings. In conclusion, our study contributes to understanding the functional analysis of the SPL gene family in Z. armatum and emphasizes the crucial role of ZaSPL21 in improving tolerance to salt and promoting flowering. The results offer potential strategies for the further utilization of these genes to improve the salt tolerance of Z. armatum.

Nitrogen (N) is a major plant nutrient and its deficiency can arrest plant growth. However, how low-N stress impair plant growth and its related tolerance mechanisms in peanut seedlings has not yet been explored. To counteract this issue, a hydroponic study was conducted to explore low N stress (0.1 mM NO₃^-) and normal (5.0 mM NO₃^-) effects on the morpho-physiological and molecular attributes of peanut seedlings. Low-N stress significantly decreased peanut plant height, leaf surface area, total root length, and primary root length after 10 days of treatment. Meanwhile, glutamate dehydrogenase, glutamine oxoglutarate aminotransferase activities, chlorophyll, and soluble protein contents were substantially decreased. Impairment in these parameters further suppressed photochemical efficiency (Fv/Fm), and chlorophyll fluorescence parameters (PI_ABS), under low-N stress. Transcriptome sequencing analysis showed a total of 2139 DEGs were identified between the two treatments. KEGG enrichment annotation analysis of DEGs revealed that 119 DEGs related to 10 pathways, including N assimilation, photosynthesis, starch, and sucrose degradation, which may respond to low-N stress in peanuts. Combined with transcriptome, small RNA, and degradome sequencing, we found that PC-3p-142756_56/A.T13EMM (CML3) and PC-5p-43940_274/A.81NSYN (YTH3) are the main modules contributing to low N stress tolerance in peanut crops. Peanut seedlings exposed to N starvation exhibited suppressed gene expression related to nitrate transport and assimilation, chlorophyll synthesis, and carbon assimilation, while also showing improved gene expression in N compensation/energy supply and carbohydrate consumption. Additionally, low N stress tolerance was strongly associated with the miRNA.

In a wake of shifting climatic scenarios, plants are frequently forced to undergo a spectrum of abiotic and biotic stresses at various stages of growth, many of which have a detrimental effect on production and survival. Naturally, microbial consortia partner up to boost plant growth and constitute a diversified ecosystem against abiotic stresses. Despite this, little is known pertaining to the interplay between endophytic microbes which release phytohormones and stimulate plant development in stressed environments. In a lab study, we demonstrated that an endophyte isolated from the Kargil region of India, a Fusarium equiseti strain K23-FE, colonizes the maize hybrid MAH 14 - 5, promoting its growth and conferring polyethylene glycol (PEG)-induced osmotic stress tolerance. To unravel the molecular mechanism, maize seedlings inoculated with endophyte were subjected to comparative transcriptomic analysis. In response to osmotic stress, genes associated with metabolic, photosynthesis, secondary metabolites, and terpene biosynthesis pathways were highly upregulated in endophyte enriched maize seedlings. Further, in a greenhouse experiment, maize plants inoculated with fungal endophyte showed higher relative leaf water content, chlorophyll content, and antioxidant enzyme activity such as polyphenol oxidase (PPO) and catalase (CAT) under 50% field capacity conditions. Osmoprotectant like proline were higher and malondialdehyde content was reduced in colonized plants. This study set as proof of concept to demonstrate that endophytes adapted to adverse environments can efficiently tweak non-host plant responses to abiotic stresses such as water deficit stress via physiological and molecular pathways, offering a huge opportunity for their deployment in sustainable agriculture.

Psa primarily utilises the type III secretion system (T3SS) to deliver effector proteins (T3Es) into host cells, thereby regulating host immune responses. However, the mechanism by which kiwifruit responds to T3SS remains unclear. To elucidate the molecular reaction of kiwifruit plants to Psa infection, M228 and mutant M228△hrcS strains were employed to inoculate Actinidia chinensis var. chinensis for performing comparative transcriptional and metabolomic analyses. Transcriptome analysis identified 973 differentially expressed genes (DEGs) related to flavonoid synthesis, pathogen interaction, and hormone signaling pathways during the critical period of Psa infection at 48 h post-inoculation. In the subsequent metabolomic analysis, flavonoid-related differential metabolites were significantly enriched after the loss of T3SS.Through multi-omics analysis, 22 differentially expressed genes related to flavonoid biosynthesis were identified. Finally, it was discovered that the transient overexpression of 3 genes significantly enhanced kiwifruit resistance to Psa. qRT-PCR analysis indicated that Ac4CL1, Ac4CL3 and AcHCT1 promote host resistance to disease, while Ac4CL3 negatively regulates host resistance to Psa. These findings enrich the plant immune regulation network involved in the interaction between kiwifruit and Psa, providing functional genes and directions with potential application for breeding kiwifruit resistance to canker disease.

Various biological processes are interconnected in plants. Transcription factors (TFs) often act as regulatory hubs to regulate plant growth and responses to stress by integrating various biological pathways. Despite extensive studies on TFs functions in various plant species, our understanding of the details of TFs regulation remains limited. In this study, clonal seedlings of Salvia miltiorrhiza were exposed to specific inhibitors for 12 h. Time-series transcriptome data, sampled hourly, were used to construct co-expression networks and gene regulatory networks (GRNs). Transcriptome dynamic analysis was utilized to capture the gene expression dynamics of various biological processes and decipher the potential molecular mechanisms that regulate these processes. The perturbation results showed the growth and development processes of S.miltiorrhiza were primarily affected at the early stage, whereas stress response-related biological processes were mainly influenced at the later stage. And there was a correlation between the series of key differentially expressed genes in terpenoid biosynthesis pathways and the topological distribution of these pathways. Furthermore, the GRNs based on TFs indicate that TFs play a crucial role in connecting various biological processes. In the cytoplasmic lysate gene regulatory module, SmWRKY48-SmTCP4-SmWRKY28 constituted a regulation hub regulating S.miltiorrhiza responses to perturbation of the MVA pathway. The regulation hub mediated various pathways, including pyruvate metabolism, glycolysis/gluconeogenesis, amino acid metabolism, and ubiquinone and other terpenoid-quinone biosynthesis.Our findings suggest that perturbation of a key biological pathway in S.miltiorrhiza has time-dependent effects on other biological processes. And SmWRKY48-SmTCP4-SmWRKY28 constitutes the regulatory hub in S.miltiorrhiza responses to perturbation of MVA pathway.