Plant Biotechnology Journal最新文献

{"title":"Precise genome editing of Dense and Erect Panicle 1 promotes rice sheath blight resistance and yield production in japonica rice","authors":"Hongyao Zhu, Tiange Zhou, Jiaming Guan, Zhuo Li, Xiurong Yang, Yuejiao Li, Jian Sun, Quan Xu, Yuan Hu Xuan","doi":"10.1111/pbi.70010","DOIUrl":"https://doi.org/10.1111/pbi.70010","url":null,"abstract":"The primary goals of crop breeding are to enhance yield and improve disease resistance. However, the “trade-off” mechanism, in which signalling pathways for resistance and yield are antagonistically regulated, poses challenges for achieving both simultaneously. Previously, we demonstrated that knock-out mutants of the <i>Dense and Erect Panicle 1</i> (<i>DEP1</i>) gene can significantly enhance rice resistance to sheath blight (ShB), and we mapped <i>DEP1</i>'s association with panicle length. In this study, we discovered that <i>dep1</i> mutants significantly reduced rice yield. Nonetheless, truncated DEP1 was able to achieve both ShB resistance and yield increase in japonica rice. To further explore the function of truncated <i>DEP1</i> in promoting yield and ShB resistance, we generated CRISPR/Cas9-mediated genome editing mutants, including a full-length deletion mutant of <i>DEP1</i>, named <i>dep1</i>, and a truncated version, <i>dep1-cys</i>. Upon inoculation with <i>Rhizoctonia solani</i>, the <i>dep1-cys</i> mutant demonstrated stronger ShB resistance than the <i>dep1</i> mutant. Additionally, <i>dep1-cys</i> increased yield per plant, whereas <i>dep1</i> reduced it. Compared to the full DEP1 protein, the truncated DEP1 (dep1-cys) demonstrated a decreased interaction affinity with IDD14 and increased affinity with IDD10, which are known to positively and negatively regulate ShB resistance through the activation of <i>PIN1a</i> and <i>ETR2</i>, respectively. The <i>dep1-cys</i> mutant exhibited higher <i>PIN1a</i> and lower <i>ETR2</i> expression than wild-type plants, suggesting that <i>dep1-cys</i> modulated IDD14 and IDD10 interactions to regulate <i>PIN1a</i> and <i>ETR2</i>, thereby enhancing ShB resistance. Overall, these data indicate that precise genome editing of <i>DEP1</i> could simultaneously improve both ShB resistance and yield, effectively mitigating trade-off regulation in rice.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"67 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143539328","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Integrative analyses reveal Bna‐miR397a–BnaLAC2 as a potential modulator of low‐temperature adaptability in Brassica napus L.","authors":"Muhammad Azhar Hussain, Yong Huang, Dan Luo, Sundas Saher Mehmood, Ali Raza, Xuekun Zhang, Yong Cheng, Hongtao Cheng, Xiling Zou, Xiaoyu Ding, Liu Zeng, Liu Duan, Bian Wu, Keming Hu, Yan Lv","doi":"10.1111/pbi.70017","DOIUrl":"https://doi.org/10.1111/pbi.70017","url":null,"abstract":"Summary<jats:italic>Brassica napus</jats:italic> L. (<jats:italic>B. napus</jats:italic>) is a major edible oil crop grown around the southern part of China, which often faces cold stress, posing potential damage to vegetative tissues. To sustain growth and reproduction, a detailed understanding of fundamental regulatory processes in <jats:italic>B. napus</jats:italic> against long‐term low temperature (LT) stress is necessary for breeders to adjust the level of LT adaption in a given region and is therefore of great economic importance. Till now, studies on microRNAs (miRNAs) in coping with LT adaption in <jats:italic>B. napus</jats:italic> are limited. Here, we performed an in‐depth analysis on two <jats:italic>B. napus</jats:italic> varieties with distinct adaptability to LT stress. Through integration of RNA sequencing (RNA‐seq) and small RNA‐sequencing (sRNA‐seq), we identified 106 modules comprising differentially expressed miRNAs and corresponding potential targets based on strong negative correlations between their dynamic expression patterns. Specifically, we demonstrated that <jats:italic>Bna‐miR397a</jats:italic> post‐transcriptionally regulates a LACCASE (LAC) gene, <jats:italic>BnaLAC2</jats:italic>, to enhance the adaption to LT stresses in <jats:italic>B. napus</jats:italic> by reducing the total lignin remodelling and ROS homeostasis. In addition, the <jats:italic>miR397</jats:italic>–<jats:italic>LAC2</jats:italic> module was also proved to improve freezing tolerance of <jats:italic>Arabidopsis</jats:italic>, indicating a conserved role of <jats:italic>miR397</jats:italic>–<jats:italic>LAC2</jats:italic> in <jats:italic>Cruciferae</jats:italic> plants. Overall, this work provides the first description of a miRNA‐mediated‐module signature for LT adaption and highlights the prominent role of laccase in future breeding programme of LT tolerant <jats:italic>B. napus</jats:italic>.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"90 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143538484","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Elucidating the enzyme network driving Amaryllidaceae alkaloids biosynthesis in Leucojum aestivum","authors":"Basanta Lamichhane, Sarah-Eve Gélinas, Natacha Merindol, Manoj Koirala, Karen Cristine Gonçalves dos Santos, Hugo Germain, Isabel Desgagné-Penix","doi":"10.1111/pbi.70026","DOIUrl":"https://doi.org/10.1111/pbi.70026","url":null,"abstract":"<i>Amaryllidaceae</i> alkaloids (AAs) are diverse bioactive metabolites with significant pharmaceutical potential, derived from 4′-O-methylnorbelladine (4′<i>O</i>M). The biosynthesis of these compounds involves the condensation of tyramine and 3,4-dihydroxybenzaldehyde by norbelladine synthase (NBS) and/or noroxomaritidine/norcraugsodine reductase (NR), followed by <i>O</i>-methylation. Cytochrome P450 enzymes, particularly the CYP96T family, introduce further structural diversity through C–C couplings, resulting in lycorine, galanthamine and crinine cores. Despite their importance, the exact biosynthetic pathways remain poorly defined. In this study, we describe key enzymes from <i>Leucojum aestivum</i> (<i>La</i>), providing crucial insight into AA biosynthesis. Transient expression in <i>Nicotiana benthamiana</i> demonstrated that <i>La</i>NBS and <i>La</i>NRII catalyse the conversion of tyramine and 3,4-dihydroxybenzaldehyde to norbelladine, which is subsequently <i>O</i>-methylated by a norbelladine-4′-<i>O</i>-methyltransferase (<i>La</i>N4′<i>O</i>MT) <i>in planta</i>. Co-agroinfiltration of <i>La</i>NBS, <i>La</i>NRII, <i>La</i>N4′<i>O</i>MT and <i>La</i>CYP96T1 resulted in the production of various phenol-coupled products, with lycorine as the predominant compound, alongside haemanthamine, crinine/vittatine and norgalanthamine. This study identifies <i>La</i>CYP96T1 and <i>La</i>CYP96T2 as the first monocot enzymes capable of catalysing all three regioselective C-C phenol couplings and also highlights the substrate promiscuity of <i>La</i>NRII. The findings not only elucidate critical steps in AA biosynthesis but also open new avenues for biotechnological application in producing valuable alkaloids, offering potential for novel drug development.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"11 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143539332","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"An efficient target-mutant screening platform of model variety Ci846 facilitates genetic studies of Setaria","authors":"Hui Zhang, Hongkai Liang, Hui Zhi, Di Yuan, Qi Yao, Renliang Zhang, Lihe Xing, Baolan Yang, Luman Sang, Lirong Zhao, Sha Tang, Liwei Wang, Hailong Wang, Yushuang Ren, Hui Zhang, Yanyan Zhang, Enbo Wang, Xinyu Man, Gongjin Xu, Linlin Zhang, Qiang He, Xianmin Diao, Guanqing Jia","doi":"10.1111/pbi.14594","DOIUrl":"https://doi.org/10.1111/pbi.14594","url":null,"abstract":"&lt;p&gt;Foxtail millet (&lt;i&gt;Setaria italica&lt;/i&gt;), one of the oldest crops originating in China, has increasingly been recognized as a promising C&lt;sub&gt;4&lt;/sub&gt; model plant due to its compact diploid genome, short growth cycle and self-pollinating nature (Li and Brutnell, &lt;span&gt;2011&lt;/span&gt;). In the past 5 years, significant breakthroughs have been achieved in its basic research and breeding, including high efficient transformation system establishment, telomere-to-telomere (T2T) genome assembly, pan-genome analysis and functional studies (He &lt;i&gt;et al&lt;/i&gt;., &lt;span&gt;2023&lt;/span&gt;; Tang &lt;i&gt;et al&lt;/i&gt;., &lt;span&gt;2023&lt;/span&gt;; Yang &lt;i&gt;et al&lt;/i&gt;., &lt;span&gt;2020&lt;/span&gt;). However, the limited genetic diversity of breeding materials and inefficiencies in identifying target mutants have continued to pose significant challenges in breeding for improved complex agronomic traits and in functional genomics research of this crop.&lt;/p&gt;\u0000&lt;p&gt;To broaden novel sources of genetic diversity and enhance mutant identification efficiency for foxtail millet improvement, we constructed a large EMS-mutagenized library and a data-sharing platform based on the model variety Ci846 (www.setariadb.com/millet/mutation, Figure S3), which is a domesticated Setaria variety with well-established transformation system and efficient indoor research platform (Wang &lt;i&gt;et al&lt;/i&gt;., &lt;span&gt;2022&lt;/span&gt;). We obtained 9243 M&lt;sub&gt;2&lt;/sub&gt; lines, of which 775 (8.38%) displayed various morphological variations under field conditions (Figure 1a,b, Table S14 and Figure S4). We sampled 4109 M&lt;sub&gt;2&lt;/sub&gt; plants and re-sequenced them using a mixed pool strategy, and obtained 1950.76 Gb of pair-end reads from 205 mixed pools, with each pool ranging from 8.6 to 12.2 Gb and an average sequencing depth of 22x (Figure 1c). This represents the largest-scale precise genotype data for mutant libraries in crop species to date. The cleaned short-read sequences were then mapped to the Yugu1-T2T reference genome. The mapping rate and coverage rate of sequencing reads were 98.21 ± 0.37% and 94.24 ± 0.37%, respectively (Table S3). A total of 2,899,449 variations were identified across the 205 mixed pools, with an average mutation density of 1/29.70 Kb across all nine chromosomes (Figure 1d). Each mixed pool contained 1,658,170 ± 11,422 SNPs and 252,784 ± 2402 indels. Among the pools, A173 had the smallest number of variations (1,875,480), while A9 had the largest number (1,989,562) (Table S4). We found that C/G to T/A nucleotide transitions were the most prevalent type (928,081, 38.7%) (Table S7) and the Thr/Val to Ala amino acid substitutions occurred at higher frequency (1.42% and 1.46%) than the average amino acid changed rate (0.16%) (Figure 1e, Table S9). Among all samples, indel size ranged from 1 to 31 bp, with an average of 12 bp, and the most frequent indel type was 1 bp deletions (104,581) (Table S5). A total of 2,368,808 mutations occurred in intergenic regions, followed by 1,850,547 upstream variants, 1,770,535 downstream vari","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"8 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-03-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143532514","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Combined enhancement of ascorbic acid, β-carotene and zeaxanthin in gene-edited lettuce","authors":"Yarin Livneh, Ehud Leor-Librach, Dor Agmon, Tal Makov-Bouaniche, Vivekanand Tiwari, Ekaterina Shor, Yelena Yeselson, Tania Masci, Arthur Schaffer, Dana Charuvi, Joseph Hirschberg, Alexander Vainstein","doi":"10.1111/pbi.70018","DOIUrl":"https://doi.org/10.1111/pbi.70018","url":null,"abstract":"Lettuce is widely grown and consumed but provides lower nutritional value compared to other leafy greens, particularly in the essential vitamins A and C. To address this, major control points in carotenoid and ascorbic acid (AsA) production were targeted using a viral-based CRISPR/Cas9 system in the commercial lettuce cultivar ‘Noga’. Knockout of <i>lycopene ε-cyclase</i> (<i>LCY-ε</i>), the enzymatic gatekeeper opposing production of β-branch carotenoids, increased β-carotene (provitamin A) levels up to 2.7-fold and facilitated zeaxanthin accumulation up to 4.3 μg/g fresh weight. Chlorophyll fluorescence measurements revealed that photosystem II efficiency was unaffected in <i>LCY-ε</i> mutants, though their non-photochemical quenching (NPQ) capacity decreased at light intensities above 400 μmol m² s^-1. However, the gene-edited plants exhibited normal growth and comparable plant mass, despite the absence of two major lettuce xanthophylls, lutein and lactucaxanthin. Modifications in a regulatory region in the upstream ORF of <i>GDP-L-galactose phosphorylase 1</i> and <i>2</i> (u<i>GGP1</i> and u<i>GGP2</i>), the rate-limiting enzyme in AsA production, resulted in an average 6.9-fold increase in AsA levels. The mutation in u<i>GGP2</i> was found to dominantly influence AsA over-accumulation. Knockout lines that combined the mutations in <i>LCY-ε</i>, u<i>GGP1</i>, u<i>GGP2</i> and in <i>carotenoid cleavage dioxygenase 4a</i> (<i>CCD4a</i>), an isozyme involved in β-carotene degradation in lettuce, exhibited significantly enhanced content of AsA, β-carotene and zeaxanthin. Our results demonstrate the potential of multi-pathway gene editing to ‘supercharge’ economically important crops such as lettuce as a means to address micronutrient deficiencies in modern diets.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"36 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-03-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143532515","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"An improved fluorescein diacetate–propidium iodide staining system for assessing microspore and pollen viability at different developmental stages","authors":"Jianian Tang, Qiyu Luo, Huali Li, Zichen Wu, Zhansheng Lin, Sensen Zhang, Letian Chen, Yao-Guang Liu","doi":"10.1111/pbi.14607","DOIUrl":"https://doi.org/10.1111/pbi.14607","url":null,"abstract":"&lt;p&gt;Fluorescein diacetate (FDA), a cell membrane-permeable esterase substrate, is widely used to assess cell viability. The esterase in viable cells catalyses hydrolysis of diacetate to produce green fluorescein (Rotman and Papermaster, &lt;span&gt;1966&lt;/span&gt;). As FDA fluorescence is positively correlated with reactive oxygen species (ROS) levels, it also is used to assess ROS biogenesis (Huang &lt;i&gt;et al&lt;/i&gt;., &lt;span&gt;2023&lt;/span&gt;). Propidium iodide (PI) is a nuclear dye that produces red fluorescence in inactivated cells, and is usually used to detect apoptosis and inactivated cells (Riccardi and Nicoletti, &lt;span&gt;2006&lt;/span&gt;). Greissl (&lt;span&gt;1989&lt;/span&gt;) first used FDA/PI staining to assess pollen viability by which viable and aborted pollen grains can be simultaneously identified. Ascari &lt;i&gt;et al&lt;/i&gt;. (&lt;span&gt;2020&lt;/span&gt;) combined FDA/PI staining with a software to automatically assess viability of mature pollen.&lt;/p&gt;\u0000&lt;p&gt;The current FDA/PI methods can only be used to analyse mature pollen because the FDA fluorescent signal produced in developing microspores is weak and quenches very quickly. The commonly used potassium iodide-iodine (I&lt;sub&gt;2&lt;/sub&gt;-KI) staining method is also suitable only for mature pollen with accumulated starch. Therefore, the current staining methods are inadequate for investigating viability and abortion processes throughout male development. The precise characterisation of pollen abortion processes usually uses cumbersome, expensive and time-consuming paraffin sectioning. Therefore, we aimed to develop an effective method to simply and efficiently detect abortion in microspores and pollen at different development stages.&lt;/p&gt;\u0000&lt;p&gt;We first used an FDA/PI solution containing higher concentrations of FDA (130 μg/mL) and PI (87 μg/mL) than previous reports, which was diluted in dH&lt;sub&gt;2&lt;/sub&gt;O as described (Jones &lt;i&gt;et al&lt;/i&gt;., &lt;span&gt;2016&lt;/span&gt;), to analyse microspore viability at tetrad stage (S8b stage) in wild-type rice (&lt;i&gt;Oryza sativa&lt;/i&gt; L.). Although FDA/PI/dH&lt;sub&gt;2&lt;/sub&gt;O-stained tetrad microspores produced green fluorescence imaged by a confocal microscopy, the green signal was quenched quickly within three minutes, followed by red fluorescence generated by PI (Figure 1a), indicating that the microspores became inactivated quickly in this solution.&lt;/p&gt;\u0000&lt;figure&gt;&lt;picture&gt;\u0000&lt;source media=\"(min-width: 1650px)\" srcset=\"/cms/asset/7e45d6ac-d16a-4f9c-bc1f-3f04d2fe6116/pbi14607-fig-0001-m.jpg\"/&gt;&lt;img alt=\"Details are in the caption following the image\" data-lg-src=\"/cms/asset/7e45d6ac-d16a-4f9c-bc1f-3f04d2fe6116/pbi14607-fig-0001-m.jpg\" loading=\"lazy\" src=\"/cms/asset/1941bb72-18d7-40ce-9845-12bde1cfc304/pbi14607-fig-0001-m.png\" title=\"Details are in the caption following the image\"/&gt;&lt;/picture&gt;&lt;figcaption&gt;\u0000&lt;div&gt;&lt;strong&gt;Figure 1&lt;span style=\"font-weight:normal\"&gt;&lt;/span&gt;&lt;/strong&gt;&lt;div&gt;Open in figure viewer&lt;i aria-hidden=\"true\"&gt;&lt;/i&gt;&lt;span&gt;PowerPoint&lt;/span&gt;&lt;/div&gt;\u0000&lt;/div&gt;\u0000&lt;div&gt;FDA/PI fluorescent staining of rice microspores and pollen. The green and r","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"31 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143517826","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Overexpression of MtNAC33 enhances biomass yield and drought tolerance in alfalfa","authors":"Ruijuan Yang, Ying Sun, Yan Zhao, Chen Bai, Yaling Liu, Jingzhe Sun, Zhaoming Wang, Feng Yuan, Xiaoshan Wang, Wenwen Liu, Chunxiang Fu","doi":"10.1111/pbi.14597","DOIUrl":"https://doi.org/10.1111/pbi.14597","url":null,"abstract":"&lt;p&gt;Alfalfa (&lt;i&gt;Medicago sativa&lt;/i&gt; L.), a highly valuable perennial forage legume, is extensively cultivated worldwide (Russelle, &lt;span&gt;2001&lt;/span&gt;). As global warming exacerbates evaporation rates, severe drought conditions, characterized by mud cracking, have increasingly affected alfalfa cultivation regions. Drought stress can decrease stomatal conductance, impair photosynthesis activity and induce reactive oxygen species (ROS) accumulation in Alfalfa plants. Therefore, it reduces alfalfa growth and accelerates flowering, leading to significant declines in biomass yield and forage quality. Previous studies have shown that the plant-specific NAC (NAM, ATAF1,2 and CUC2) transcription factors play crucial roles in plant response to diverse environmental stresses. For example, NACs are involved in cold response of tomato, salt tolerance of soybean and disease resistance of &lt;i&gt;Arabidopsis&lt;/i&gt;. Recent studies have also highlighted that OsNAC120 and OsNAC016 regulated the balance between plant growth and drought tolerance by promoting gibberellin (GA) biosynthesis, brassinosteroid (BR) signalling and repressing abscisic acid (ABA)-mediated drought responses (Wu &lt;i&gt;et al&lt;/i&gt;., &lt;span&gt;2022&lt;/span&gt;; Xie &lt;i&gt;et al&lt;/i&gt;., &lt;span&gt;2024&lt;/span&gt;). These insights provide a framework for developing crop varieties with improved biomass yield under drought conditions.&lt;/p&gt;\u0000&lt;p&gt;The &lt;i&gt;Medicago truncatula&lt;/i&gt; NAC transcription factor MtNAC33 (Medtr3g096140), one member of the NAC2 subfamily, clusters phylogenetically with &lt;i&gt;Arabidopsis&lt;/i&gt; NAC082 and NAC103 (Figure S1). Previous studies revealed that &lt;i&gt;MtNAC33&lt;/i&gt; is induced by mannitol and NaCl treatments in &lt;i&gt;Medicago&lt;/i&gt; seedlings (Ling &lt;i&gt;et al&lt;/i&gt;., &lt;span&gt;2017&lt;/span&gt;), but its biological functions remain largely unexplored. To elucidate the role of &lt;i&gt;MtNAC33&lt;/i&gt; in drought tolerance, its expression was analysed under mannitol-simulated drought and NaCl-induced salt stress. Results confirmed significant induction of &lt;i&gt;MtNAC33&lt;/i&gt; expression in these stress conditions (Figure 1a; Figure S2). To assess its functional role, &lt;i&gt;MtNAC33&lt;/i&gt; was overexpressed in &lt;i&gt;Arabidopsis thaliana&lt;/i&gt;. Two transgenic lines, MtNAC33OE-A and MtNAC33OE-C, with the highest &lt;i&gt;MtNAC33&lt;/i&gt; expression levels, were selected for analysis. Under drought stress (10 days without watering), MtNAC33OE plants exhibited enhanced drought resistance (Figure S3) but showed no significant differences with wild-type plants under salt stress (Figure S4).&lt;/p&gt;\u0000&lt;figure&gt;&lt;picture&gt;\u0000&lt;source media=\"(min-width: 1650px)\" srcset=\"/cms/asset/df058da8-84ac-4d32-b519-67950b50e458/pbi14597-fig-0001-m.jpg\"/&gt;&lt;img alt=\"Details are in the caption following the image\" data-lg-src=\"/cms/asset/df058da8-84ac-4d32-b519-67950b50e458/pbi14597-fig-0001-m.jpg\" loading=\"lazy\" src=\"/cms/asset/a97debe3-c7b9-48e4-b492-e0ce7b3352ce/pbi14597-fig-0001-m.png\" title=\"Details are in the caption following the image\"/&gt;&lt;/picture&gt;&lt;figcaption&gt;\u0000&lt;div&gt;&lt;strong&gt;Figure 1&lt;span style=\"font-weight:normal","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"22 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143517827","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Naturally occurring spinach defensins confer tolerance to citrus greening and potato zebra chip diseases","authors":"Carmen S. Padilla, Sonia C. Irigoyen, Manikandan Ramasamy, Mona B. Damaj, Michelle M. Dominguez, Denise Rossi, Renesh H. Bedre, William O. Dawson, Choaa El-Mohtar, Michael S. Irey, Kranthi K. Mandadi","doi":"10.1111/pbi.70013","DOIUrl":"https://doi.org/10.1111/pbi.70013","url":null,"abstract":"&lt;p&gt;Citrus greening or Huanglongbing (HLB) and potato zebra chip (ZC) are devastating crop diseases worldwide (Mora &lt;i&gt;et al&lt;/i&gt;., &lt;span&gt;2021&lt;/span&gt;; Stelinski &lt;i&gt;et al&lt;/i&gt;., &lt;span&gt;2024&lt;/span&gt;). The diseases are associated with two related, fastidious (unculturable), phloem-limited bacteria, ‘&lt;i&gt;Candidatus&lt;/i&gt; Liberibacter asiaticus’ (&lt;i&gt;C&lt;/i&gt;Las) and ‘&lt;i&gt;Ca&lt;/i&gt;. Liberibacter solanacearum’ (&lt;i&gt;C&lt;/i&gt;Lso) that occurs in the United States. They are transmitted by the insect vector &lt;i&gt;Diaphorina citri&lt;/i&gt; Kuwayama and &lt;i&gt;Bactericera cockerelli&lt;/i&gt; (Sulc.), respectively (Mora &lt;i&gt;et al&lt;/i&gt;., &lt;span&gt;2021&lt;/span&gt;).&lt;/p&gt;\u0000&lt;p&gt;Defensins are short (~40 to 50 amino acids) basic, cysteine-rich peptides integral to the innate immune system in plants, animals, and insects and possess broad-spectrum inhibitory activity against bacterial and fungal pathogens (Cornet &lt;i&gt;et al&lt;/i&gt;., &lt;span&gt;1995&lt;/span&gt;; Velivelli &lt;i&gt;et al&lt;/i&gt;., &lt;span&gt;2018&lt;/span&gt;). Here, we evaluated whether overexpressing defensins from spinach in citrus and potato can confer tolerance to ‘&lt;i&gt;Ca&lt;/i&gt;. Liberibacter spp.’ diseases.&lt;/p&gt;\u0000&lt;p&gt;First, we characterized defensin-encoding genes from spinach (&lt;i&gt;Spinacia oleracea&lt;/i&gt;) (Mirkov and Mandadi, &lt;span&gt;2020&lt;/span&gt;; Segura &lt;i&gt;et al&lt;/i&gt;., &lt;span&gt;1998&lt;/span&gt;). The spinach defensins (&lt;i&gt;So&lt;/i&gt;AMPs) are evolutionarily closer to Group II defensins of &lt;i&gt;Arabidopsis&lt;/i&gt;, rice and &lt;i&gt;Medicago&lt;/i&gt; (Figure S1a). They possess the conserved Gamma-thionin/knottin-fold and multiple cysteine residues in the amino acid sequence (Figure S1b), and three characteristic antiparallel &lt;i&gt;β&lt;/i&gt;-sheets and an &lt;i&gt;α&lt;/i&gt;-helix, stabilized by disulfide bridges in the predicted ternary structure (Figure S1c) (Cornet &lt;i&gt;et al&lt;/i&gt;., &lt;span&gt;1995&lt;/span&gt;).&lt;/p&gt;\u0000&lt;p&gt;Next, we evaluated the efficacy of spinach defensins spp. using &lt;i&gt;Rhizobium rhizogenes&lt;/i&gt;-mediated hairy root transformation (Irigoyen &lt;i&gt;et al&lt;/i&gt;., &lt;span&gt;2020&lt;/span&gt;). Transgene expression was driven under the &lt;i&gt;Cauliflower mosaic virus&lt;/i&gt; (CaMV) 35S promoter in the ‘&lt;i&gt;Ca&lt;/i&gt;. Liberibacter spp.’ infected hairy roots (Figure 1a) (Irigoyen &lt;i&gt;et al&lt;/i&gt;., &lt;span&gt;2020&lt;/span&gt;). Both &lt;i&gt;So&lt;/i&gt;AMP1 and &lt;i&gt;So&lt;/i&gt;AMP2 expressing hairy roots showed 71–99% reduction (&lt;i&gt;P&lt;/i&gt; ≤ 0.05 or &lt;i&gt;P&lt;/i&gt; ≤ 0.01) of ‘&lt;i&gt;Ca&lt;/i&gt;. Liberibacter spp.’ compared to negative controls (empty vector) (Figure 1b) (Table S1). Next, stable potato transgenic lines expressing &lt;i&gt;SoA&lt;/i&gt;MP1 and &lt;i&gt;So&lt;/i&gt;AMP2 were generated using the &lt;i&gt;Agrobacterium tumefaciens&lt;/i&gt;-mediated plant transformation. Two independent transgenic lines and non-transformed (NT) plants (negative controls) were challenged with &lt;i&gt;C&lt;/i&gt;Lso-carrying potato psyllids in controlled no-choice assays. The non-transformed plants developed characteristic zebra chip-associated shoot chlorosis and yellowing symptoms at 28 days post-infection (Figure 1c). Strikingly, the &lt;i&gt;So&lt;/i&gt;AMP-expressing transgenic plants showed attenuated disease symptoms (Figure 1c), reduced &lt;i&gt;C&lt;/i&gt;Lso titre (2.1–5.2% for &lt;i&gt;So&lt;/i","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"31 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143517828","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Calcium-dependent protein kinase PpCDPK29-mediated Ca2+-ROS signal and PpHSFA2a phosphorylation regulate postharvest chilling tolerance of peach fruit","authors":"Liangyi Zhao, Hua Cassan-Wang, Yaqin Zhao, Yinqiu Bao, Yuanyuan Hou, Yu Liu, Zhengguo Wu, Mondher Bouzayen, Yonghua Zheng, Peng Jin","doi":"10.1111/pbi.70024","DOIUrl":"https://doi.org/10.1111/pbi.70024","url":null,"abstract":"Green and chemical-free hot water (HW) treatment can effectively reduce the chilling injury of peach fruit; however, the mechanism of inducing chilling resistance by heat treatment is still unclear. This study found that HW treatment could activate reactive oxygen species (ROS) signalling, forming ROS-Ca²⁺ signalling. Furthermore, we identified a peach Ca²⁺ sensor, calcium-dependent protein kinase 29 (PpCDPK29), as a positive regulator of postharvest chilling resistance. PpCDPK29 interacted with ROS-generating proteins (PpRBOHC/D) and antioxidant enzymes (PpSOD and PpCAT1) to jointly maintain ROS homeostasis. Meanwhile, we found that PpHSFA2a was phosphorylated by PpCDPK29 and transferred to the nucleus, which enhanced the binding ability of PpHSFA2a to the target genes. Here, PpHSFA2a activated the transcription of target genes <i>PpHSP18.5</i>, <i>PpHSP70</i>, <i>PpGSTU7</i>, <i>PpGSTU19</i>, <i>PpGolS1</i> and <i>PpBAM1</i>, acted as molecular chaperones, improved ROS scavenging and enhanced osmoregulation to alleviate postharvest chilling injury of peach fruit. In summary, HW treatment could alleviate postharvest chilling injury in peach fruit by activating the PpCDPK29-mediated Ca²⁺-ROS and HSF-HSP signalling pathways, providing a novel signalling network for postharvest quality control of peach fruit.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"51 1","pages":""},"PeriodicalIF":13.8,"publicationDate":"2025-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143517825","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Harnessing promoter elements to enhance gene editing in plants: perspectives and advances.","authors":"Nikita Gondalia, Luis Felipe Quiroz, Linyi Lai, Avinash Kumar Singh, Moman Khan, Galina Brychkova, Peter C McKeown, Manash Chatterjee, Charles Spillane","doi":"10.1111/pbi.14533","DOIUrl":"https://doi.org/10.1111/pbi.14533","url":null,"abstract":"<p><p>Genome-edited plants, endowed with climate-smart traits, have been promoted as tools for strengthening resilience against climate change. Successful plant gene editing (GE) requires precise regulation of the GE machinery, a process controlled by the promoters, which drives its transcription through interactions with transcription factors (TFs) and RNA polymerase. While constitutive promoters are extensively used in GE constructs, their limitations highlight the need for alternative approaches. This review emphasizes the promise of tissue/organ specific as well as inducible promoters, which enable targeted GE in a spatiotemporal manner with no effects on other tissues. Advances in synthetic biology have paved the way for the creation of synthetic promoters, offering refined control over gene expression and augmenting the potential of plant GE. The integration of these novel promoters with synthetic systems presents significant opportunities for precise and conditional genome editing. Moreover, the advent of bioinformatic tools and artificial intelligence is revolutionizing the characterization of regulatory elements, enhancing our understanding of their roles in plants. Thus, this review provides novel insights into the strategic use of promoters and promoter editing to enhance the precision, efficiency and specificity of plant GE, setting the stage for innovative crop improvement strategies.</p>","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":" ","pages":""},"PeriodicalIF":10.1,"publicationDate":"2025-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143514043","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}