Plant Biotechnology Reports最新文献

Long non-coding RNAs (lncRNAs) are transcripts longer than 200 nucleotides that lack significant protein coding potential and have been shown to regulate various biological processes. This study was designed to identify lncRNAs in coconut and their role in the process of somatic embryogenesis in coconut, a crop with high recalcitrance to in vitro culture. RNA-Seq data of coconut embryogenic calli of the West Coast Tall cultivar was exploited for in silico prediction of lncRNA. From a total of 6328 transcripts, which were annotated as uncharacterised or with no homology hits with the existing database, 5110 putative lncRNAs are identified. We also studied the relationship between lncRNAs, microRNAs (miRNAs) and mRNAs and found that some of the lncRNAs act as miRNA precursors, some as potential miRNA targets and some function as endogenous target mimics (eTMs) for miRNAs. Real-time quantitative PCR confirmed that 10 selected lncRNAs showed significant differences in the expression pattern in different stages of coconut somatic embryogenesis. Our results suggest the existence of diverse lncRNAs in coconut embryogenic calli, some of which are differentially expressed. The information generated in this study could be of great value in understanding the molecular mechanisms governing somatic embryogenesis in coconut.

Abstract

Expansin plays a crucial role in plant growth and stress resistance as a cell wall relaxation protein. The expansin family consists of four subfamilies: EXPA, EXPB, EXLA, and EXLB. However, a few reports have been previously published investigating EXLA genes. The research here aimed to characterize the PtEXLA1 gene from a popular species (P. alba × P. glandulosa CV.84K) and evaluate its role through genetic transformation to understand its contribution to plant growth and stress resistance. The results showed that the PtEXLA1 gene was 780 bp in length, encoded 259 amino acids, and had typical characteristics of EXLA. The PtEXLA1 transgenic tobacco plants had a larger corolla in comparison to wild-type plants, and exhibited higher resistance to drought, high temperature, and salt stress based on the evaluation of chlorophyll content, relative conductivity, and malondialdehyde content. PtEXLA1 can be an efficient gene resource for stress resistance breeding of plants.

The ATP-binding cassette (ABC) transporter family is one of the largest protein families in plants and plays an essential role in addressing biotic and abiotic stresses. Wheat, a vital global grain crop, faces multifaceted safety challenges, primarily from fungal diseases like stripe rust and powdery mildew. In the present study, we identified the whole genome of the wheat ABC family, and 463 nonredundant ABC genes were identified. The ABC family can be divided into nine evolutionary branches and eight subfamilies based on phylogenetic tree analysis. This paper delved deeper into characterizing the gene structure, promoter region, and gene expression within the TaABC family. Segmental duplication was the main reason for the expansion of the TaABC genes. Ka/Ks analysis suggested that most TaABC genes were intensely purified and selected. The collinear analysis of TaABC and other species showed that the ABC genes were conserved in evolution. RNA-seq data and qPCR data from wheat infected with powdery mildew or stripe rust showed that most TaABC genes were induced to change expression. The candidate genes TaABCB15-3B and TaABCG38 exhibited responsiveness to powdery mildew in resistant/susceptible wheat, while remaining unresponsive to stripe rust. Our findings serve as a valuable reference for gaining a deeper understanding of the function and evolution of TaABCs, aiding in the identification of enduring disease resistance genes within the TaABCs of wheat.

The performance of crop plants is critically affected by biotic and abiotic stress. These stressors threaten food availability by reducing overall crop yield and productivity. Changes in chromatin state by epigenetic modification are part of plant adaptive and survival responses and are considered pivotal for improving agronomic traits. Epigenetics is an exciting field that involves heritable gene expression changes that do not require changes in DNA sequence. Epigenetic modification is well known as a crucial player in plant phenotypic diversity and defense against pathogens. Hence, there is a growing interest in unlocking the epigenome for crop improvement. Herein, we highlight the epigenetic modifications implicated in plant biotic stress response and their contributions to important agronomic traits. We also discussed adopting epigenetics to expand phenotypic diversity and produce desired characteristics in crop plants.

Floral architecture plays a pivotal role in developmental processes under genetic regulation and is also influenced by environmental cues. This affects the plant silique phenotype in Arabidopsis and grain yield in crops. Despite the relatively small number of family members of zinc finger homeodomain (ZF-HD) transcription factors (TFs) in plants, their biological role needs to be investigated to understand the molecular mechanisms associated with plant developmental processes. Therefore, we generated HB31SRDX and HB21SRDX repressor mutant lines to understand the functional role of ZF-HD TFs. The mutant lines showed severe defects in plant architecture, including increased branching number, reduced plant height, distorted floral phenotype, and short silique. We found that HB31 and HB21 are paralogs in Arabidopsis, and both positively regulate cell size-related genes, cell wall modification factor-related genes, and M-type MADS-box TF families. In addition, HB31 and HB21 are negatively associated with abiotic stress-related genes, vegetative-to-reproductive phase transition of meristem-related genes, and TCP and RAV TFs. microRNA164 (miR164), miR822, miR396, miR2934, and miR172 were downregulated, whereas miR169, miR398, miR399, and miR157 were upregulated in the two repressor lines. Phenotypic and molecular analyses demonstrated that the miR396/GRF modules regulated by HB31 and HB21 are involved in the plan floral architecture of Arabidopsis. The findings of this study will help elucidate the role of ZF-HD TFs in maintaining the floral architecture.

Pesticides have been an integral part of modern agriculture as their use ensures good harvests. However, excessive use of pesticides in the last few decades has caused significant environmental degradation. Moreover, excessive use of pesticides causes stress on crops and non-target plants and exhibits toxicity to other organisms including mammals, microbes, and insects. Plants employ various morphological, physiological, and biochemical mechanisms to reduce pesticides toxicity. One such mechanism is production of secondary metabolites that improves stress tolerance of plants. In addition, recent studies have also highlighted a potential role of plasma technology in mitigating various abiotic and biotic environmental stresses. Besides, plasma treatment improves seed germination, physiological processes, and seedling establishment during the early growth stages of a plant under adverse and non-adverse conditions and thus can be used an alternate to the pesticide treatment. This review article summarizes recent advancements in understanding the synthesis, accumulation, and transportation of secondary metabolites which have significant relevance to crop improvement programs. We also present an overview of the effects of plasma treatment on phytopathogenic bacterial cell suspensions and plant responses to metabolic activity. In the future, researchers need to develop innovative ideas to reduce the use of chemical pesticides in farming practices.

Six popular and widely cultivated arabica coffee (Coffea arabica L.) varieties of commercial importance namely Selection 5B, Selection 13, Selection 11, Selection 8, Selection 7.3 and Selection 3 were tested for their genetic identity with ISSR markers. Fifteen ISSR primers were tested using genomic DNA of selected coffee varieties. Pooled genomic DNA of all the six varieties was amplified with each ISSR primer with an average of four loci per primer. The size range of locus amplified by all the fifteen primers was ranging from 100 to 1200 bp depending upon on the ISSR primers. Only three out of fifteen primers, namely ISSR4, ISSR6 and ISSR8, were screened based on the number of amplified locus and size range from low to high. The selective ISSR primers distinguished all the six varieties of coffee with unique markers. ISSR 4 amplified two unique markers with a locus size 1300 bp and 950 bp for Sln.5B and 180 bp and 150 bp for Sln.13. ISSR6 had produced five varietal-specific markers with a locus size of 180 bp in Sln.5B, 1250 bp in Sln.11, 350 bp in Sln.3. ISSR8 had amplified seven unique loci across the coffee varieties with 700 bp and 800 bp in Sln.5B, 200 bp and 500 bp in Sln.11 and one locus each in Sln.7.3 and Sln.3 with 300 bp and 150 bp respectively. Repeated amplification of genomic DNA of all the six varieties of coffee with selective ISSR primers produced consistent ISSR genetic fingerprints. Selective ISSR primers were validated with marker parameters resolving power (RP), effective multiplex ratio (EMR), marker index (MI) and polymorphic information content (PIC). Utilisation of these markers in arabica coffee genetic improvement is discussed.

Multiple myeloma (MM) is an incurable disease characterized by malignant plasma cells within the bone marrow, and its increasing occurrence has highlighted the need for innovative strategies to address relapse and treatment resistance. Given the substantial expression of programmed death ligand 1 (PD-L1) in the human multiple myeloma cell line RPMI8226, we propose PD-L1 as a promising target for multiple myeloma therapy. Here, we successfully engineered an anti-PD-L1 monoclonal antibody (mAb) within a plant-based system. Building upon our previous findings, we germinated seeds derived from transgenic plants under in vitro conditions. Afterward, we screened the resulting seedlings for expression of the anti-PD-L1 mAb using polymerase chain reaction (PCR) and western blot analyses. Anti-PD-L1 mAbs were successfully purified from plant leaves and characterized through SDS-PAGE analysis. Our findings, which were confirmed via indirect enzyme-linked immunosorbent assay (ELISA), validate the binding affinity of the anti-PD-L1 mAb to recombinant PD-L1 protein. Furthermore, we investigated the interaction between the plant-derived anti-PD-L1 mAb and Fc gamma receptor I (FcγRI) as well as Fc gamma receptor IIIa (FcγRIIIa) molecules, confirming robust affinity. Additionally, the antibody’s binding affinity to the human multiple myeloma cancer cell line RPMI8226 was confirmed via cell ELISA. Our findings demonstrated that, unlike existing therapeutics, the plant-derived anti-PD-L1 antibody not only effectively binds to human recombinant PD-L1 protein but also to FcγRI and FcγRIIIa. These findings suggest the potential of plant-derived anti-PD-L1 mAb for the development of innovative therapies against multiple myeloma, emphasizing the need for further research and preclinical evaluation.

Brassica is an essential genus in agriculture, and gene flow from living modified (LM) organisms to native relatives or non-modified cultivars of the crucifer family is a critical issue in Asia. Loop-mediated isothermal amplification (LAMP) is a simple, rapid, and accurate method for identifying LM crops. This study aimed to develop a LAMP assay for the laboratory diagnosis and field analysis of three novel LM canola events: T45, 73496, and MON88302. The genomic DNA of each LM canola was amplified by incubating at 63 °C for 30 min with a newly developed 2 × LAMP premix containing Bst DNA polymerase and event-specific primer sets. The sensitivity of the LAMP assay for the three canola events in our condition ranged from 100 pg·μL⁻¹ to 1 ng·μL⁻¹. Genetic elements of the three LM events were identified through a combination of the LAMP assay, a syringe-type DNA extraction kit, and a portable isothermal amplifier. This study proposes a novel, simple system based on isothermal amplification for the DNA detection of the three LM canola events in a survey field.

Photosynthesis is responsible for sustained plant productivity and ensures food supply. The change in global climatic patterns affects photosynthesis that subsequently reduces plant yield and poses threat to food security. Photosynthesis relies on a dual nature enzyme ribulose 1, 5 bisphosphate carboxylase oxygenase (Rubisco), which can fix CO₂ as well as O₂. The fixation rate of CO₂ to O₂ depends upon the relative concentration of CO₂ inside chloroplast. Higher level of CO₂ results in improved photosynthesis, however, its concentration depends upon environmental conditions. Under adverse climate conditions, the CO₂ level drops down that leads to increased oxygenation which impedes the photosynthesis and reduces plant productivity. The impact is more significant and apparent specifically in C₃ plants. Attempts have been made to address the loss in photosynthesis and multiple strategies have been adapted to date that focus on improvement of photosynthesis in C₃ plants. In this review, we have discussed the multiple strategies being employed by different researchers to date for improvement of photosynthesis. The strategies discussed in this review include: improving the performance of Rubisco, engineering CO₂-concentrating mechanism of C₄ photosynthesis into C₃ species, transformation of bicarbonate transporters from cyanobacteria into chloroplasts of C₃ plants, and establishment of photorespiratory bypasses to catabolise toxic glycolate in shortest possible pathway.