{"title":"A simple, cost-effective, and efficient method for screening CRISPR/Cas9 mutants in plants","authors":"","doi":"10.1016/j.jplph.2024.154375","DOIUrl":"10.1016/j.jplph.2024.154375","url":null,"abstract":"<div><div>The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9)-mediated genome editing system is widely used for targeted mutagenesis in a growing number of plant species. To streamline the screening process for mutants, especially those generated from low-efficiency editing events, there is a need for a rapid, cost-effective, and efficient method. Although several screening methods have been developed to process initial samples, these methods often tend to be time-consuming, expensive, or inefficient when dealing with larger sample sizes. Here we describe a simple, rapid, low-cost, and sensitive screening method for screening CRISPR/Cas9 mutants called PCR-<em>Bsl</em> I-associated analysis (PCR-BAA). This method requires only standard PCR and <em>Bsl</em> I restriction enzyme digestion, as well as agarose gel electrophoresis analysis. This method is particularly well suited for the efficient screening of mutants from larger populations of transformants. The simplicity, low cost, and high sensitivity of the PCR-BAA method make it particularly suitable for rapid screening of CRISPR/Cas9-induced mutants, especially those from low-efficiency editing events.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":null,"pages":null},"PeriodicalIF":4.0,"publicationDate":"2024-10-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142586492","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Alkaline tolerance in plants: The AT1 gene and beyond","authors":"","doi":"10.1016/j.jplph.2024.154373","DOIUrl":"10.1016/j.jplph.2024.154373","url":null,"abstract":"<div><div>Salt stress poses a serious challenge to crop production and a significant threat to global food security and ecosystem sustainability. Soil salinization commonly occurs in conjunction with alkalization, which causes combined saline–alkaline stress. Alkaline soil predominantly comprises NaHCO<sub>3</sub> and Na<sub>2</sub>CO<sub>3</sub> and is characterized by a high pH. The combined saline–alkaline stress is more harmful to crop production than neutral salt stress owing to the effects of both elevated salinity and high pH stress. Through genome association analysis of sorghum, a recent study has identified <em>Alkaline tolerance 1</em> (<em>AT1</em>) as a contributor to alkaline sensitivity in crops. <em>AT1</em>, which is the first gene to be identified as being specifically associated with alkaline tolerance, encodes a G protein γ-subunit (Gγ). Editing of <em>AT1</em> enhances the yields of sorghum, rice, maize, and millet grown in alkaline soils, indicating that <em>AT1</em> has potential for generating alkaline-resistant crops. In this review, we summarize the role of <em>AT1</em> in alkaline tolerance in plants and present a phylogenetic analysis along with a motif comparison of Gγ subunits of monocot and dicot plants across various species.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":null,"pages":null},"PeriodicalIF":4.0,"publicationDate":"2024-10-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142502616","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Developmental-specific regulation promotes the free amino acids accumulation in chlorotic tea plants (Camellia sinensis)","authors":"","doi":"10.1016/j.jplph.2024.154371","DOIUrl":"10.1016/j.jplph.2024.154371","url":null,"abstract":"<div><div>Chlorophyll-deficient tea plant exhibits a significantly higher accumulation of free amino acids (FAAs) than normal tea plants. This study focused on the impact of leaf color and the developmental stage on FAAs in six tea germplasms while maintaining all other conditions. The total FAAs content initially increased as the leaf matured during the one-bud-two-leaves (1B2L) and one-bud-three-leaves (1B3L) stages in green germplasms, then decreased or stabilized in the one-bud-four-leaves (1B4L) stage. In contrast, chlorotic germplasms showed continuous FAAs' content increase from 1B2L to 1B4L, thus being significantly positively correlated with total chlorophyll content. Interestingly, ethylamine content decreased with leaf maturation in both chlorotic and green germplasms, thus showing a significant negative correlation with L-theanine content only in chlorotic germplasms. Comparative RNA-seq analysis linked FAAs accumulation in chlorotic germplasm's 1B3L to photosynthesis inhibition and in 1B4L to nitrogen assimilation promotion. Feeding experiments revealed higher L-theanine synthesis and degradation abilities in chlorotic shoots versus green shoots, with synthesis efficiency exceeding degradation efficiency. Overall, this study uncovers a developmental-specific FAAs accumulation pattern in chlorotic germplasms and offers novel insights into the precise regulation by leaf color and developmental stage.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":null,"pages":null},"PeriodicalIF":4.0,"publicationDate":"2024-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142502617","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"SPOTLIGHT: Orchestrating nitrogen metabolism: Histone modifications and its interplay with the biological clock","authors":"","doi":"10.1016/j.jplph.2024.154369","DOIUrl":"10.1016/j.jplph.2024.154369","url":null,"abstract":"","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":null,"pages":null},"PeriodicalIF":4.0,"publicationDate":"2024-10-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142441842","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Adaptive mechanisms of wheat cultivars to lead toxicity through enhanced oxidative defense, ionomic redistribution, and anatomical modifications","authors":"","doi":"10.1016/j.jplph.2024.154370","DOIUrl":"10.1016/j.jplph.2024.154370","url":null,"abstract":"<div><div>Lead (Pb) contamination is a critical environmental issue that poses a substantial threat to agricultural sustainability and crop productivity, particularly for staple crops like wheat (<em>Triticum aestivum</em> L.). This study investigates the differential physiological, biochemical, and anatomical responses of two wheat cultivars, SKD-1 and Borlaug-16, under Pb stress (100 mg/kg Pb for 21 days). Borlaug-16 displayed a notable tolerance to Pb toxicity, evidenced by a significant increase in total biomass, including a 41.22% rise in shoot turgid weight and a 23.37% increase in root turgid weight, alongside a 57.72% enhancement in root cortex thickness. This cultivar also showed increased antioxidant enzyme activities, such as catalase and peroxidase, and a better ionomic balance, maintaining higher levels of essential minerals like Ca in leaf tissues while effectively accumulating Pb and other trace elements in roots. In contrast, SKD-1 suffered from a more substantial reduction in essential minerals and weaker anatomical and biochemical defenses. The study's novelty lies in providing an integrated approach to understanding wheat cultivar-specific adaptations to Pb stress, suggesting Borlaug-16 as a promising candidate for cultivation in Pb-contaminated soils. These findings underscore the importance of developing Pb-tolerant cultivars to ensure sustainable wheat production in polluted environments.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":null,"pages":null},"PeriodicalIF":4.0,"publicationDate":"2024-10-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142468330","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Tripartite interactions between grapevine, viruses, and arbuscular mycorrhizal fungi provide insights into modulation of oxidative stress responses","authors":"","doi":"10.1016/j.jplph.2024.154372","DOIUrl":"10.1016/j.jplph.2024.154372","url":null,"abstract":"<div><div>Arbuscular mycorrhizal fungi (AMF) can be beneficial for plants exposed to abiotic and biotic stressors. Although widely present in agroecosystems, AMF influence on crop responses to virus infection is underexplored, particularly in woody plant species such as grapevine. Here, a two-year greenhouse experiment was set up to test the hypothesis that AMF alleviate virus-induced oxidative stress in grapevine. The ‘Merlot’ cultivar was infected with three grapevine-associated viruses and subsequently colonized with two AMF inocula, containing one or three species, respectively. Five and fifteen months after AMF inoculation, lipid peroxidation - LPO as an indicator of oxidative stress and indicators of antioxidative response (proline, ascorbate - AsA, superoxide dismutase - SOD, ascorbate- APX and guaiacol peroxidases - GPOD, polyphenol oxidase - PPO, glutathione reductase - GR) were analysed. Expression of genes coding for a stilbene synthase (<em>STS1</em>), an enhanced disease susceptibility (<em>EDS1</em>) and a lipoxygenase (<em>LOX</em>) were determined in the second harvesting. AMF induced reduction of AsA and SOD over both years, which, combined with not AMF-triggered APX and GR, suggests decreased activation of the ascorbate-glutathione cycle. In the mature phase of the AM symbiosis establishment GPOD emerged as an important mechanism for scavenging H<sub>2</sub>O<sub>2</sub> accumulation. These results, together with reduction in <em>STS1</em> and increase in <em>EDS1</em> gene expression, suggest more efficient reactive oxygen species scavenging in plants inoculated with AMF. Composition of AMF inocula was important for proline accumulation. Overall, our study improves the knowledge on ubiquitous grapevine-virus-AMF systems in the field, highlighting that established functional AM symbiosis could reduce virus-induced stress.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":null,"pages":null},"PeriodicalIF":4.0,"publicationDate":"2024-10-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142445933","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Crosstalk of methylglyoxal and calcium signaling in maize (Zea mays L.) thermotolerance through methylglyoxal-scavenging system","authors":"","doi":"10.1016/j.jplph.2024.154362","DOIUrl":"10.1016/j.jplph.2024.154362","url":null,"abstract":"<div><div>Methylglyoxal (MG) and calcium ion (Ca<sup>2+</sup>) can increase multiple-stress tolerance including plant thermotolerance. However, whether crosstalk of MG and Ca<sup>2+</sup> exists in the formation of maize thermotolerance and underlying mechanism still remain elusive. In this paper, maize seedlings were irrigated with MG and calcium chloride alone or in combination, and then exposed to heat stress (HS). The results manifested that, compared with the survival percentage (SP, 45.3%) of the control seedlings, the SP of MG and Ca<sup>2+</sup> alone or in combination was increased to 72.4%, 74.2%, and 83.4% under HS conditions, indicating that Ca<sup>2+</sup> and MG alone or in combination could upraise seedling thermotolerance. Also, the MG-upraised SP was separately weakened to 42.2%, 40.3%, 52.1%, and 39.4% by Ca<sup>2+</sup> chelator (ethylene glycol tetraacetic acid, EGTA), plasma membrane Ca<sup>2+</sup> channel blocker (lanthanum chloride, LaCl<sub>3</sub>), intracellular Ca<sup>2+</sup> channel blocker (neomycin, NEC), and calmodulin (CaM) antagonist (trifluoperazine, TFP). However, significant effect of MG scavengers N-acetylcysteine (NAC) and aminoguanidine (AG) on Ca<sup>2+</sup>-induced thermotolerance was not observed. Similarly, an endogenous Ca<sup>2+</sup> level in seedlings was increased by exogenous MG under non-HS and HS conditions, while exogenous Ca<sup>2+</sup> had no significant effect on endogenous MG. These data implied that Ca<sup>2+</sup> signaling, at least partly, mediated MG-upraised thermotolerance in maize seedlings. Moreover, the activity and gene expression of glyoxalase system (glyoxalase I, glyoxalase II, and glyoxalase III) and non-glyoxalase system (MG reductase, aldehyde reductase, aldo-keto reductase, and lactate dehydrogenase) were up-regulated to a certain extent by Ca<sup>2+</sup> and MG alone in seedlings under non-HS and HS conditions. The up-regulated MG-scavenging system by MG was enhanced by Ca<sup>2+</sup>, while impaired by EGTA, LaCl<sub>3</sub>, NEC, or TFP. These data suggest that the crosstalk of MG and Ca<sup>2+</sup> signaling in maize thermotolerance through MG-scavenging system. These findings provided a theoretical basis for breeding climate-resilient maize crop and developing smart agriculture.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":null,"pages":null},"PeriodicalIF":4.0,"publicationDate":"2024-10-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142424402","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Involvement of plasma membrane H+-ATPase in the nitrate-nutrition uptake and utilization in indica rice","authors":"","doi":"10.1016/j.jplph.2024.154368","DOIUrl":"10.1016/j.jplph.2024.154368","url":null,"abstract":"<div><div>Utilization of nitrogen by crops is essential for sustainable agriculture. The transport of nitrate (NO<sub>3</sub><sup>−</sup>) across the plasma membrane is a critical gateway for N uptake and subsequent utilization. This process requires proton (H<sup>+</sup>) coupled cotransport, which is driven by proton motive force, provided by plasma membrane (PM) H<sup>+</sup>-ATPase. In this report, two indica rice varieties [Meixiangzhan 2 (MXZ) and Jifengyou 1002 (JFY)] in South China were selected and cultivated in hydroponic solution with 0.5 mM or 2.0 mM NO<sub>3</sub><sup>−</sup> as the N source. The JFY exhibited stronger growth with higher biomass than MXZ under both 0.5 mM and 2.0 mM NO<sub>3</sub><sup>−</sup>. PM H<sup>+</sup>-ATPase activity of JFY roots was significantly higher than that of MXZ. The higher PM H<sup>+</sup>-ATPase activity in JFY was consistent with a higher abundance of PM H<sup>+</sup>-ATPase protein and higher transcription levels of <em>OSAs</em>, such as <em>OSA2</em>, <em>OSA7</em> and <em>OSA8</em> in roots, <em>OSA3</em>, <em>OSA7</em> and <em>OSA8</em> in leaves. The expression of nitrate transporters (<em>OsNRT1;1b</em>, <em>OsNRT2.1</em>, <em>OsNRT2.2</em>, and <em>OsNAR2.1</em>) were also higher in roots or shoots of JFY than those in MXZ. Under 0.5 mM and 2.0 mM NO<sub>3</sub><sup>−</sup>, the NO<sub>3</sub><sup>−</sup> absorption and translocation rate, nitrate content, as well as nitrate reductase (NR) activity were all significantly higher in JFY, as compared to those in MXZ. Taken together, in JFY and MXZ, a higher level of PM H<sup>+</sup>-ATPase protein and higher activity coupled with greater efficiency in nitrate uptake, translocation and assimilation, suggesting the existence of a close correlation between PM H<sup>+</sup>-ATPase and nitrate utilization in indica rice. PM H<sup>+</sup>-ATPase may one of the elite genes that can contribute to nitrate use efficiency in rice.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":null,"pages":null},"PeriodicalIF":4.0,"publicationDate":"2024-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142406530","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"A mycoheterotrophic orchid uses very limited soil inorganic nitrogen in its natural habitat","authors":"","doi":"10.1016/j.jplph.2024.154367","DOIUrl":"10.1016/j.jplph.2024.154367","url":null,"abstract":"<div><div>Mycoheterotrophic plants acquire nitrogen (N) directly from the soil and through their symbiotic fungi. The fungi-derived N has received considerable attention, but the contribution of soil-derived N has been largely overlooked. We investigated how the leafless, rootless, and almost mycoheterotrophic orchid <em>Cymbidium macrorhizon</em> obtains soil N by applying <sup>15</sup>N-labeled ammonium nitrate in its natural habitat, and tracking metabolite accumulation and mycorrhizal fungal association after N application. The decline of N in the rhizome from flowering to fruiting indicated a transfer of N from the rhizome to fruits. At current dose of N application (0.6 g NH<sub>4</sub>NO<sub>3</sub> each plant), only 1.5% of the plant's N was derived from fertilizer, resulting in a low nitrogen use efficiency of 0.27%. The majority of those newly absorbed N (88.89%) was found sank in the rhizome. Amino acids (or their derivatives) and alkaloids were predominant differentially accumulated nitrogenous metabolites after N application, with amino acids occurring in both fruits and the rhizome, and alkaloids primarily in the fruits. The addition of N did not alter the richness of mycorrhizal fungi, but did affect their relative abundance. Our findings suggest that <em>Cymbidium macrorhizon</em> uses very limited soil inorganic nitrogen in its natural habitat, and the root-like rhizome primarily stores N rather than absorbs its inorganic forms, offering new insights into how mycoheterotrophic plants utilize soil N, and the influence of nutrient availability on the orchid-fungi association.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":null,"pages":null},"PeriodicalIF":4.0,"publicationDate":"2024-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142381152","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Phlorotannins contribute to the ameliorative bioactivities of Ecklonia maxima-derived bioproduct in salt-stressed Solanum lycopersicum","authors":"","doi":"10.1016/j.jplph.2024.154366","DOIUrl":"10.1016/j.jplph.2024.154366","url":null,"abstract":"<div><div>Seaweed-derived bioproducts are increasingly being deployed as an environmentally friendly and sustainable approach to crop management under stressful growth conditions including salinity. The bioactivities of seaweed-derived extracts are linked to the presence of diverse groups of bioactive compounds. In the present study, the phlorotannins present in the seaweed <em>Ecklonia maxima</em> and Kelpak®, an <em>E. maxima</em>-derived bioproduct, were quantified and identified. Three phlorotannins were identified in <em>E. maxima</em>, namely eckol, 2-phloroeckol, and dibenzodioxin-fucodiphloroethol. Eckol (589.11 – 822.54 μg l<sup>−1</sup>) and dibenzodioxin-fucodiphloroethol (85 – 895 μg l<sup>−1</sup>) were present in Kelpak®. Phlorotannin bioactivity was investigated in tomato seedlings grown under NaCl-induced salinity stress. The seedlings treated with either individual phlorotannins (i.e., eckol or a fraction containing 2-phloroeckol and dibenzodioxin-fucodiphloroethol) or Kelpak® resulted in a reprogramming of biomass allocation as indicated by an increased root-to-shoot ratio. Phlorotannin and Kelpak® treatments induced the accumulation of antioxidants with an attendant augmentation of the antioxidant capacities and inhibition of membrane damage in the NaCl-stressed seedlings. Kelpak® treatment induced an increase in abscisic acid (ABA) accumulation and phlorotannin treatments lowered the ABA content of the stressed seedlings. These results demonstrated that phlorotannins contributed to the ameliorative actions of Kelpak®. The more potent effects of Kelpak®, in comparison to phlorotannins, in improving dry matter accumulation, ABA content, antioxidative properties, and inhibiting tissue injury of the salt-stressed tomato seedlings may be attributed to the presence of other bioactive components in the Kelpak® product.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":null,"pages":null},"PeriodicalIF":4.0,"publicationDate":"2024-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142391383","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}