Hydrogen sulfide (H2S) and its mediated protein persulfidation regulate fruit ripening. Fruit aroma, a key quality trait formed during ripening, largely determines consumer preferences. However, the mechanism by which H2S modulates aroma biosynthesis in pear remains unclear. This study reveals that H2S inhibited aroma biosynthesis in ‘Nanguo’ pear and downregulated the fatty acid desaturase gene PuFAD8. Transient overexpression and silencing assays confirmed that PuFAD8 was required for linoleic acid synthesis, a key aroma precursor. Heterologous overexpression of PuFAD8 in tomato and its knockout in tomato further validated that PuFAD8 promoted fruit aroma formation, while H2S suppressed this function. Transcriptome analysis identified transcription factors PuMYB21 and PuERF106 co-expressed with PuFAD8. Dual-luciferase and yeast one-hybrid assays revealed that PuMYB21 bound and activated the PuFAD8 promoter. Moreover, Y2H, luciferase complementation, pull-down, and transient expression assays demonstrated that PuERF106 interacted with PuMYB21, synergistically enhancing aroma biosynthesis. Persulfidation at Cys162 of PuERF106 attenuated its interaction with PuMYB21, thereby inhibiting aromatic compound synthesis in pear fruit. Subsequently, the biological function of the PuERF106-PuMYB21-PuFAD8 molecular module was validated using stable transgenic pear callus lines and a panel of heterologous and endogenous transgenic tomato lines. This study reveals a novel regulatory mechanism where H2S-mediated persulfidation suppresses pear aroma biosynthesis, providing a theoretical basis for genetic improvement of high-quality pear.
{"title":"Hydrogen sulfide-mediated persulfidation of ERF106 controls pear fruit aroma biosynthesis via disrupting the ERF106-MYB21-FAD8 complex","authors":"Zhuoyan Xu, Kangdi Hu, Longfei Yan, Shuwei Wei, Weisheng Zhao, Huihui Song, Gaifang Yao, Hua Zhang","doi":"10.1111/tpj.71129","DOIUrl":"10.1111/tpj.71129","url":null,"abstract":"<p>Hydrogen sulfide (H<sub>2</sub>S) and its mediated protein persulfidation regulate fruit ripening. Fruit aroma, a key quality trait formed during ripening, largely determines consumer preferences. However, the mechanism by which H<sub>2</sub>S modulates aroma biosynthesis in pear remains unclear. This study reveals that H<sub>2</sub>S inhibited aroma biosynthesis in ‘Nanguo’ pear and downregulated the fatty acid desaturase gene <i>PuFAD8</i>. Transient overexpression and silencing assays confirmed that <i>PuFAD8</i> was required for linoleic acid synthesis, a key aroma precursor. Heterologous overexpression of PuFAD8 in tomato and its knockout in tomato further validated that <i>PuFAD8</i> promoted fruit aroma formation, while H<sub>2</sub>S suppressed this function. Transcriptome analysis identified transcription factors PuMYB21 and PuERF106 co-expressed with <i>PuFAD8</i>. Dual-luciferase and yeast one-hybrid assays revealed that PuMYB21 bound and activated the <i>PuFAD8</i> promoter. Moreover, Y2H, luciferase complementation, pull-down, and transient expression assays demonstrated that PuERF106 interacted with PuMYB21, synergistically enhancing aroma biosynthesis. Persulfidation at Cys162 of PuERF106 attenuated its interaction with PuMYB21, thereby inhibiting aromatic compound synthesis in pear fruit. Subsequently, the biological function of the PuERF106-PuMYB21-PuFAD8 molecular module was validated using stable transgenic pear callus lines and a panel of heterologous and endogenous transgenic tomato lines. This study reveals a novel regulatory mechanism where H<sub>2</sub>S-mediated persulfidation suppresses pear aroma biosynthesis, providing a theoretical basis for genetic improvement of high-quality pear.</p>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"127 6","pages":""},"PeriodicalIF":6.2,"publicationDate":"2026-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/tpj.71129","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148987333","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Kun Li, Jiangtao Jia, Yong Yang, Minghui Zhang, Shulin Zhang, Xiaohan Liang, Zhiwen Lu, Min Xu, Zhe Wang, Fengbo Yang, Haipeng Li, Jinggong Guo, Kun-peng Jia, Lam-Son Phan Tran, Jose R. Botella, Weiqiang Li, Yuchen Miao
Drought stress has a profound impact on yield, with massive repercussions for agricultural production. In addition to ABA, the central hormone controlling the plant response to drought, secondary metabolites also play important functions in drought stress; however, the precise nature of their roles is still obscure. Metabolomics analysis of ABA- and PEG-treated Arabidopsis wild–type (WT) seedlings as well as ABA signal transduction mutants snrk-triple (snrk2.2 snrk2.3 snrk2.6) and pyl-quadruple (pyr1 pyl1 pyl2 pyl4) identified a number of drought-related metabolites. Specifically, we observed differences in the levels of sinapic acid and quercetin. To assess the function of sinapoyl esters in the regulation of drought stress, we characterized the sinapoylglucose accumulator 1 (sng1) mutant, deficient in the conversion of sinapoylglucose (SG) to sinapoylmalate (SM), resulting in high endogenous levels of SG and low SM levels. sng1 mutants showed increased drought resistance compared to WT plants. RNA-sequencing of sng1 and WT rosette leaves before and after dehydration revealed differentially expressed genes related to cuticle synthesis, anthocyanin biosynthesis, leaf senescence and ABA response. Consistently, phenotypic analysis of the sng1 mutant showed increased cuticle synthesis, wax crystal deposition, anthocyanin accumulation, leaf senescence, ABA responsiveness and stomatal closure compared to WT plants. We propose that the deficiency of SM and accumulation of SG in sng1 mutants may affect the metabolic flow of sinapoyl esters to activate multiple biological processes whose combined action enhances drought resistance. Our findings shed light on the secondary metabolites regulatory network in response to drought stress in Arabidopsis.
{"title":"Sinapoylglucose: malate sinapoyltransferase SNG1 negatively regulates drought resistance by crosstalk with abscisic acid signaling in Arabidopsis","authors":"Kun Li, Jiangtao Jia, Yong Yang, Minghui Zhang, Shulin Zhang, Xiaohan Liang, Zhiwen Lu, Min Xu, Zhe Wang, Fengbo Yang, Haipeng Li, Jinggong Guo, Kun-peng Jia, Lam-Son Phan Tran, Jose R. Botella, Weiqiang Li, Yuchen Miao","doi":"10.1111/tpj.71102","DOIUrl":"10.1111/tpj.71102","url":null,"abstract":"<p>Drought stress has a profound impact on yield, with massive repercussions for agricultural production. In addition to ABA, the central hormone controlling the plant response to drought, secondary metabolites also play important functions in drought stress; however, the precise nature of their roles is still obscure. Metabolomics analysis of ABA- and PEG-treated Arabidopsis wild–type (WT) seedlings as well as ABA signal transduction mutants <i>snrk-triple</i> (<i>snrk2.2 snrk2.3 snrk2.6</i>) and <i>pyl-quadruple</i> (<i>pyr1 pyl1 pyl2 pyl4</i>) identified a number of drought-related metabolites. Specifically, we observed differences in the levels of sinapic acid and quercetin. To assess the function of sinapoyl esters in the regulation of drought stress, we characterized the <i>sinapoylglucose accumulator 1</i> (<i>sng1</i>) mutant, deficient in the conversion of sinapoylglucose (SG) to sinapoylmalate (SM), resulting in high endogenous levels of SG and low SM levels. <i>sng1</i> mutants showed increased drought resistance compared to WT plants. RNA-sequencing of <i>sng1</i> and WT rosette leaves before and after dehydration revealed differentially expressed genes related to cuticle synthesis, anthocyanin biosynthesis, leaf senescence and ABA response. Consistently, phenotypic analysis of the <i>sng1</i> mutant showed increased cuticle synthesis, wax crystal deposition, anthocyanin accumulation, leaf senescence, ABA responsiveness and stomatal closure compared to WT plants. We propose that the deficiency of SM and accumulation of SG in <i>sng1</i> mutants may affect the metabolic flow of sinapoyl esters to activate multiple biological processes whose combined action enhances drought resistance. Our findings shed light on the secondary metabolites regulatory network in response to drought stress in Arabidopsis.</p>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"127 5","pages":""},"PeriodicalIF":6.2,"publicationDate":"2026-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/tpj.71102","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148987318","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
<p>For years, Zm00001d033286 sat in the maize reference genome annotated as a phenylalanine ammonia-lyase (PAL). It was a reasonable designation since the enzyme genuinely has PAL activity. Here, Sakamoto and colleagues show that this annotation captures only one part of its chemistry (Sakamoto et al., <span>2026</span>). The enzyme can also move the amino group of tyrosine from the α- to the β-carbon, producing the non-proteinogenic amino acid (R)-β-tyrosine. Renamed ZmTAM1, it combines tyrosine aminomutase (TAM), PAL and tyrosine ammonia-lyase (TAL) activities on a single scaffold. Its product inhibits the radicle growth of <i>Striga hermonthica</i>, a parasitic weed responsible for severe losses in maize production.</p><p>For Sakamoto, the discovery reinforced ‘the importance of experimentally verifying gene function rather than relying solely on database annotations’. The rice enzyme OsTAM1 was also originally catalogued as a PAL (Yan et al., <span>2015</span>), leading him to suspect that ‘there are more examples like this waiting to be discovered’.</p><p>PAL is a gateway between primary metabolism and phenylpropanoid biosynthesis (Zhang & Liu, <span>2015</span>). In grasses, many PAL enzymes also accept tyrosine and are therefore designated phenylalanine/TALs, or PTALs (Barros et al., <span>2016</span>). PAL and TAL reactions release a substrate's amino group as ammonia. Aminomutases use the same MIO (4-methylideneimidazole-5-one) cofactor but relocate the amino group to the β-carbon instead (Figure 1). Although TAM had long been known in bacteria, it was identified in plants only in 2015 (Christenson et al., <span>2003</span>; Yan et al., <span>2015</span>).</p><p>Screening 22 grass species, the authors detected appreciable β-tyrosine in only two: oat, at 49 nmol g<sup>−1</sup> fresh weight, and maize, at 39 nmol g<sup>−1</sup>. Wheat, barley and sorghum contained barely a trace. Because oat and maize belong to the distantly related BOP and PACMAD clades, respectively, β-tyrosine production appears scattered across the grass family rather than confined to a single lineage.</p><p>The evidence for aminomutase chemistry in maize is elegantly direct. The authors fed maize seedlings tyrosine carrying heavy carbon and nitrogen labels and recovered both labels in the β-tyrosine produced. The signal for the product retaining the heavy nitrogen was 3.6 times that for the corresponding carbon-labelled product lacking it. A pathway in which tyrosine was first deaminated and later reaminated would have lost the original nitrogen; its retention therefore strongly supports a direct TAM route.</p><p>A homology search identified Zm00001d033286, itself annotated as a PAL, as the maize protein most similar to OsTAM1, at 88% amino acid identity. It was also the only maize PAL or PTAL to carry tyrosine and asparagine at positions Y128 and N449, corresponding to the Y125 and N446 residues known to be important for TAM activity in the rice enzyme. Transient
多年来，Zm00001d033286在玉米参考基因组中被标记为苯丙氨酸解氨酶（PAL）。这是一个合理的名称，因为这种酶确实具有PAL活性。在这里，Sakamoto及其同事表明，该注释仅捕获了其化学成分的一部分（Sakamoto et al., 2026）。该酶还能将酪氨酸的氨基从α-移到β-碳上，生成非蛋白氨基酸(R)-β-酪氨酸。它被重新命名为ZmTAM1，在一个支架上结合了酪氨酸氨基交换酶（TAM）、PAL和酪氨酸氨解酶（TAL）的活性。它的产品抑制了玉米生产中严重损失的寄生杂草Striga hermonthica的根生长。对于坂本来说，这一发现强调了“通过实验验证基因功能的重要性，而不是仅仅依靠数据库注释”。水稻酶OsTAM1最初也被归类为PAL (Yan et al., 2015)，这让他怀疑“还有更多这样的例子等待被发现”。PAL是初级代谢和苯丙类生物合成之间的通道（Zhang & Liu, 2015）。在草中，许多PAL酶也接受酪氨酸，因此被称为苯丙氨酸/ tal或PTALs （Barros等，2016）。PAL和TAL反应释放底物的氨基为氨。氨基转氨酶使用相同的MIO（4-甲基咪唑-5- 1）辅助因子，但将氨基转移到β-碳上（图1）。虽然早在细菌中就发现了TAM，但直到2015年才在植物中发现（Christenson et al., 2003; Yan et al., 2015）。在筛选22种牧草时，作者仅在两种牧草中检测到显著的β-酪氨酸：燕麦（鲜重为49 nmol g−1）和玉米（鲜重为39 nmol g−1）。小麦、大麦和高粱几乎没有。由于燕麦和玉米分别属于亲缘关系较远的BOP和PACMAD分支，β-酪氨酸的产生似乎分散在整个草科，而不是局限于一个单一的谱系。玉米中氨基转氨酶化学的证据非常直接。作者给玉米幼苗喂带有重碳和重氮标签的酪氨酸，并在产生的β-酪氨酸中回收了这两种标签。保留重氮的产品的信号是缺少重氮的相应碳标记产品的3.6倍。酪氨酸首先脱氨基，然后再氨基的途径会失去原来的氮；因此，它的保留强烈支持直接TAM路线。同源性研究发现，Zm00001d033286是与OsTAM1最相似的玉米蛋白，氨基酸同源性为88%。它也是唯一在Y128和N449位点携带酪氨酸和天冬酰胺的玉米PAL或PTAL，对应于已知在水稻酶中对TAM活性重要的Y125和N446残基。在烟叶中的瞬时表达产生(R)-β-酪氨酸，并伴有在玉米提取物中发现的少量(S)成分，其中(R)-形式的对映体过量为98%。因此，作者将该蛋白重新命名为ZmTAM1。用PTALs中发现的组氨酸取代Y128会急剧减少β-酪氨酸的产生，而用相应的赖氨酸取代N449则会消除β-酪氨酸的产生，MIO结合基序的破坏也会消除β-酪氨酸的产生。用纯化酶进行的体外实验得出了最惊人的结果：ZmTAM1同时接受酪氨酸和苯丙氨酸，并表现出TAM、TAL和PAL活性。TAM反应的催化效率最高，其次是PAL和TAL，而在实验条件下未检测到苯丙氨酸氨基转氨酶活性。那么，为什么这种酶重新定位了酪氨酸的氨基，却释放了苯丙氨酸的氨基？根据对高粱PTAL的研究（Jun et al., 2018）， Sakamoto提出苯丙氨酸遵循弗里德尔-克拉茨型路线，其中氨从活性位点逃逸。相比之下，酪氨酸可能会遵循E2路线，将氮保留足够长的时间，以便转移到β-碳上。这种机制上的相容性可以解释一个支架是如何支持三种反应的：正如Sakamoto所说，氨基转化酶化学“可以被视为氨解酶反应的延伸”。系统发育将TAM谱系置于PAL而不是PTAL分支中，这表明在PTALs和TAM中使用酪氨酸的能力是独立进化的。PAL和TAM谱系之间的分裂可能早于BOP和PACMAD草的分化，尽管它们的共同祖先是否已经具有TAM活性仍未解决。没有短柄草、小麦、大麦、芒草或谷子的序列属于TAM分支，这种模式与重复的基因丢失相一致。Sakamoto认为这可能反映了一种代谢权衡：芳香氨基酸的生产成本很高，所以TAM可能在β-酪氨酸的好处“没有超过它的代谢成本”的情况下失去了。然而，单靠序列并不能提供全部答案。 高粱蛋白Sobic.001G160500和水稻蛋白LOC_Os11g48110位于TAM分支内，携带相同的诊断性Tyr和Asn残基。然而，高粱不积累可观的β-酪氨酸，并且LOC_Os11g48110蛋白先前被证明缺乏TAM活性（Yan et al., 2015）。因此，这两个残基很重要，但它们本身是不够的，留下其他的结构特征，表达模式或两者都有待鉴定。β-酪氨酸可能的生物学功能同样令人着迷。在实验室生物测定中，40 μM (R)-β-酪氨酸使S. hermonthica根伸长减少17% （Sakamoto等，2026）。这个浓度接近茉莉酸处理过的玉米根的估计，尽管在实际感染期间存在的量仍然未知。此外，玉米没有向培养基中分泌可检测到的β-酪氨酸，这使得附着前的保护不太可能。因此，重要的测试将是该化合物是否在吸器形成期间或之后影响斯特里加，寄生虫通过吸器渗透其宿主。燕麦可能是最诱人的未解决问题。这是调查中最高的β-酪氨酸积累体，但在现有燕麦资源中未发现携带诊断残基的序列。Sakamoto怀疑燕麦酶“可能仍然没有被注释，这可以解释为什么氨基酸序列搜索没有检测到它”。代谢物大量存在；导致这种疾病的基因仍有待分类。在应用方面，Sakamoto认为育种比将β-酪氨酸作为外部化合物应用更有前景，特别是因为玉米已经产生了它。他仍然保持谨慎：单独增加β-酪氨酸可能不能提供完全的保护，它对活玉米植株感染的影响尚未得到证实。然而，加强现有的化学防御可以为抵抗寄生杂草提供新的途径。因此，由于注释的限制，本文在开始的地方结束了。目前在PAL家族中发现了两种植物tam，它们都隐藏在PAL标签后面，而燕麦代谢数据表明至少还有一种未被注释。这些基因可能已经存在于参考基因组中；认识到他们实际在做什么需要实验。
{"title":"Hidden in plain sight: a maize ‘PAL’ with aminomutase activity","authors":"Charlay Wood","doi":"10.1111/tpj.71122","DOIUrl":"10.1111/tpj.71122","url":null,"abstract":"<p>For years, Zm00001d033286 sat in the maize reference genome annotated as a phenylalanine ammonia-lyase (PAL). It was a reasonable designation since the enzyme genuinely has PAL activity. Here, Sakamoto and colleagues show that this annotation captures only one part of its chemistry (Sakamoto et al., <span>2026</span>). The enzyme can also move the amino group of tyrosine from the α- to the β-carbon, producing the non-proteinogenic amino acid (R)-β-tyrosine. Renamed ZmTAM1, it combines tyrosine aminomutase (TAM), PAL and tyrosine ammonia-lyase (TAL) activities on a single scaffold. Its product inhibits the radicle growth of <i>Striga hermonthica</i>, a parasitic weed responsible for severe losses in maize production.</p><p>For Sakamoto, the discovery reinforced ‘the importance of experimentally verifying gene function rather than relying solely on database annotations’. The rice enzyme OsTAM1 was also originally catalogued as a PAL (Yan et al., <span>2015</span>), leading him to suspect that ‘there are more examples like this waiting to be discovered’.</p><p>PAL is a gateway between primary metabolism and phenylpropanoid biosynthesis (Zhang & Liu, <span>2015</span>). In grasses, many PAL enzymes also accept tyrosine and are therefore designated phenylalanine/TALs, or PTALs (Barros et al., <span>2016</span>). PAL and TAL reactions release a substrate's amino group as ammonia. Aminomutases use the same MIO (4-methylideneimidazole-5-one) cofactor but relocate the amino group to the β-carbon instead (Figure 1). Although TAM had long been known in bacteria, it was identified in plants only in 2015 (Christenson et al., <span>2003</span>; Yan et al., <span>2015</span>).</p><p>Screening 22 grass species, the authors detected appreciable β-tyrosine in only two: oat, at 49 nmol g<sup>−1</sup> fresh weight, and maize, at 39 nmol g<sup>−1</sup>. Wheat, barley and sorghum contained barely a trace. Because oat and maize belong to the distantly related BOP and PACMAD clades, respectively, β-tyrosine production appears scattered across the grass family rather than confined to a single lineage.</p><p>The evidence for aminomutase chemistry in maize is elegantly direct. The authors fed maize seedlings tyrosine carrying heavy carbon and nitrogen labels and recovered both labels in the β-tyrosine produced. The signal for the product retaining the heavy nitrogen was 3.6 times that for the corresponding carbon-labelled product lacking it. A pathway in which tyrosine was first deaminated and later reaminated would have lost the original nitrogen; its retention therefore strongly supports a direct TAM route.</p><p>A homology search identified Zm00001d033286, itself annotated as a PAL, as the maize protein most similar to OsTAM1, at 88% amino acid identity. It was also the only maize PAL or PTAL to carry tyrosine and asparagine at positions Y128 and N449, corresponding to the Y125 and N446 residues known to be important for TAM activity in the rice enzyme. Transient","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"127 5","pages":""},"PeriodicalIF":6.2,"publicationDate":"2026-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/tpj.71122","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148987347","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The application of Green Revolution alleles Rht-B1b and Rht-D1b in wheat semi-dwarf breeding has remarkably improved lodging resistance and increased global wheat production. However, these gibberellin (GA)-insensitive genes require higher water and fertilizer inputs and impair seedling emergence and early vigor, prompting the search for alternative GA-sensitive dwarfing genes to breed environmentally friendly wheat varieties for sustainable wheat production. Here, we reported the map-based cloning of Rht4 and functional characterization of its potential causal gene TaBSK3-2B, which encodes a plasma membrane-localized serine/threonine-protein kinase involved in the brassinosteroid (BR) signaling pathway. Loss function of TaBSK3-2B attenuated BR responses and inhibited stem elongation by reducing cell expansion, which was accompanied by decreased levels of the bioactive gibberellins GA1 and GA4. Under controlled conditions, TaBSK3-2B positively regulates wheat drought tolerance by integrating transcriptional level changes of key drought-related genes and physiological adjustments under water deficit. Furthermore, we identified a natural haplotype TaBSK3-2BH2 which was associated with reduced plant height and improved yield-related traits, with the potential to mitigate the yield penalty associated with the original Rht4 allele, a major limitation on its application in wheat breeding. The frequency of TaBSK3-2BH2 is dramatically higher in cultivars compared to landraces, indicating a strong artificial selection of TaBSK3-2BH2 during semi-dwarf wheat breeding. Our study not only provides new insights into the synergistic improvement of plant height and grain yield by fine-tuning BR signaling but also offers potential genetic resources for breeding drought-adapted semi-dwarf wheat varieties.
{"title":"Deletion of TaBSK3-2B attenuates BR signaling to regulate the Rht4 semi-dwarf phenotype and drought tolerance in wheat","authors":"Qiumei Lu, Shan Lu, Zining Wang, Yihan Men, Renmei Tian, Yue Qiao, Xianglan Kong, Zhangchen Zhao, Chunge Cui, Yanju Huang, Liang Chen, Yin-gang Hu, Linzhou Huang","doi":"10.1111/tpj.71118","DOIUrl":"10.1111/tpj.71118","url":null,"abstract":"<p>The application of Green Revolution alleles <i>Rht-B1b</i> and <i>Rht-D1b</i> in wheat semi-dwarf breeding has remarkably improved lodging resistance and increased global wheat production. However, these gibberellin (GA)-insensitive genes require higher water and fertilizer inputs and impair seedling emergence and early vigor, prompting the search for alternative GA-sensitive dwarfing genes to breed environmentally friendly wheat varieties for sustainable wheat production. Here, we reported the map-based cloning of <i>Rht4</i> and functional characterization of its potential causal gene <i>TaBSK3-2B</i>, which encodes a plasma membrane-localized serine/threonine-protein kinase involved in the brassinosteroid (BR) signaling pathway. Loss function of <i>TaBSK3-2B</i> attenuated BR responses and inhibited stem elongation by reducing cell expansion, which was accompanied by decreased levels of the bioactive gibberellins GA<sub>1</sub> and GA<sub>4</sub>. Under controlled conditions, <i>TaBSK3-2B</i> positively regulates wheat drought tolerance by integrating transcriptional level changes of key drought-related genes and physiological adjustments under water deficit. Furthermore, we identified a natural haplotype <i>TaBSK3-2B</i><sup><i>H2</i></sup> which was associated with reduced plant height and improved yield-related traits, with the potential to mitigate the yield penalty associated with the original <i>Rht4</i> allele, a major limitation on its application in wheat breeding. The frequency of <i>TaBSK3-2B</i><sup><i>H2</i></sup> is dramatically higher in cultivars compared to landraces, indicating a strong artificial selection of <i>TaBSK3-2B</i><sup><i>H2</i></sup> during semi-dwarf wheat breeding. Our study not only provides new insights into the synergistic improvement of plant height and grain yield by fine-tuning BR signaling but also offers potential genetic resources for breeding drought-adapted semi-dwarf wheat varieties.</p>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"127 5","pages":""},"PeriodicalIF":6.2,"publicationDate":"2026-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/tpj.71118","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148981908","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Shun-Ling Tan, Antoni M. Vera-Vives, Heyu Wang, Xing Huang, Claudia Beraldo, Alessandro Alboresi, Tomas Morosinotto
Mitochondrial respiration catalyses the transfer of electrons from NADH to oxygen through the activity of five multiprotein complexes. In plants, mitochondria also possess additional alternative electron transport pathways, including the alternative oxidase (AOX) pathway, which transfers electrons from ubiquinol to O2, bypassing the cytochrome-dependent electron transport chain. This study investigates the functional role of AOX during plant evolution by analysing Physcomitrium patens plants that lack or overexpress AOX. In the moss P. patens, AOX exhibits a remarkably high electron transport capacity, sufficient to fully compensate for the inactivation of the cytochrome pathway. Despite the high potential activity of AOX, aox knockout lines did not show significant defects in growth under various abiotic stresses. This suggests that the cytochrome pathway can compensate for AOX loss and effectively support the consumption of reducing power produced by photosynthesis even under stressful conditions. Although photosynthetically active cells can produce ATP in chloroplasts under illumination independently of mitochondrial electron transport, respiration is shown to be essential for the conversion of reducing power into ATP for distribution throughout the cell. Simultaneous inactivation of AOX and CIII was lethal, indicating that AOX and the complementary cytochrome pathway contribute to the essential role of mitochondrial respiration in the redox and energy balance in P. patens cells.
{"title":"Essential mitochondrial activity in Physcomitrium patens relies on complementary cytochrome and alternative oxidase pathways","authors":"Shun-Ling Tan, Antoni M. Vera-Vives, Heyu Wang, Xing Huang, Claudia Beraldo, Alessandro Alboresi, Tomas Morosinotto","doi":"10.1111/tpj.71127","DOIUrl":"10.1111/tpj.71127","url":null,"abstract":"<p>Mitochondrial respiration catalyses the transfer of electrons from NADH to oxygen through the activity of five multiprotein complexes. In plants, mitochondria also possess additional alternative electron transport pathways, including the alternative oxidase (AOX) pathway, which transfers electrons from ubiquinol to O<sub>2</sub>, bypassing the cytochrome-dependent electron transport chain. This study investigates the functional role of AOX during plant evolution by analysing <i>Physcomitrium patens</i> plants that lack or overexpress AOX. In the moss <i>P. patens,</i> AOX exhibits a remarkably high electron transport capacity, sufficient to fully compensate for the inactivation of the cytochrome pathway. Despite the high potential activity of AOX, <i>aox</i> knockout lines did not show significant defects in growth under various abiotic stresses. This suggests that the cytochrome pathway can compensate for AOX loss and effectively support the consumption of reducing power produced by photosynthesis even under stressful conditions. Although photosynthetically active cells can produce ATP in chloroplasts under illumination independently of mitochondrial electron transport, respiration is shown to be essential for the conversion of reducing power into ATP for distribution throughout the cell. Simultaneous inactivation of AOX and CIII was lethal, indicating that AOX and the complementary cytochrome pathway contribute to the essential role of mitochondrial respiration in the redox and energy balance in <i>P. patens</i> cells.</p>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"127 5","pages":""},"PeriodicalIF":6.2,"publicationDate":"2026-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/tpj.71127","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148981832","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Zikai Zhou, Di Miao, Jie Zhao, Yuxi Zhou, Ting Yan, Niu Niu, Zhuo Bao, Fangjiao Lv, Lu Liu, Ning Xu, Fang Yan, Jun Liu, Hada Wuriyanghan
The typical nucleotide-binding leucine-rich repeat (NLR) proteins, such as TIR-NBS-LRR (TNL) and CC-NBS-LRR (CNL), are known to be engaged in effector-triggered immunity (ETI). However, several atypical resistance (R) proteins with truncated NLR domains or with nonclassical domains are emerging to be key regulators in plant immunity. SMV resistance cluster 4 (SRC4) is an atypical NLR protein in soybean, distinguished by an N-terminal Domain (NTD)-TIR-shortened NBS oligomerization region (SNOR)-EF-hand (EFh) domain architecture. SRC4 lacks both NBS and LRR domains but retains basal antiviral activity through its TIR-SNOR region, with SNOR contributing to TIR-dependent antiviral activity and SRC4 self-association. The EFh domain recognizes coat protein (CP) of soybean mosaic virus (SMV) and serves as a Ca2+-responsive module to distinctly regulate TIR-SNOR-mediated immune response in soybean. SRC4-overexpression (Ox-SRC4) soybean plants show SMV resistance without obvious growth penalty. Furthermore, several SRC4 high-expression soybean varieties showed stronger SMV resistance and Ca2+ responsiveness than the SRC4 low-expression soybean varieties. Our study highlights the essential roles of atypical resistance proteins in plant immunity and growth-defense balance.
典型的核苷酸结合富亮氨酸重复序列（NLR）蛋白，如TIR-NBS-LRR （TNL）和CC-NBS-LRR (CNL)，已知参与效应触发免疫（ETI）。然而，一些具有截断NLR结构域或具有非经典结构域的非典型抗性(R)蛋白正在成为植物免疫的关键调节因子。SMV抗性簇4 （SRC4）是大豆非典型NLR蛋白，具有n端结构域(NTD)- tir -缩短的NBS寡聚区(SNOR)-EF-hand （EFh）结构域结构。SRC4缺乏NBS和LRR结构域，但通过其TIR-SNOR区域保持基本的抗病毒活性，SNOR有助于tir依赖性抗病毒活性和SRC4的自结合。EFh结构域识别大豆花叶病毒（SMV）外壳蛋白（CP），并作为Ca2+响应模块明显调节tir - snorr介导的大豆免疫应答。src4过表达（Ox-SRC4）的大豆植株表现出SMV抗性，但没有明显的生长损失。此外，一些SRC4高表达大豆品种比SRC4低表达大豆品种表现出更强的SMV抗性和Ca2+响应性。我们的研究强调了非典型抗性蛋白在植物免疫和生长防御平衡中的重要作用。
{"title":"An atypical EF-hand domain containing resistance protein SRC4 boosts plant immunity in a calcium-enhanced manner","authors":"Zikai Zhou, Di Miao, Jie Zhao, Yuxi Zhou, Ting Yan, Niu Niu, Zhuo Bao, Fangjiao Lv, Lu Liu, Ning Xu, Fang Yan, Jun Liu, Hada Wuriyanghan","doi":"10.1111/tpj.71125","DOIUrl":"10.1111/tpj.71125","url":null,"abstract":"<p>The typical nucleotide-binding leucine-rich repeat (NLR) proteins, such as TIR-NBS-LRR (TNL) and CC-NBS-LRR (CNL), are known to be engaged in effector-triggered immunity (ETI). However, several atypical resistance (R) proteins with truncated NLR domains or with nonclassical domains are emerging to be key regulators in plant immunity. SMV resistance cluster 4 (SRC4) is an atypical NLR protein in soybean, distinguished by an N-terminal Domain (NTD)-TIR-shortened NBS oligomerization region (SNOR)-EF-hand (EFh) domain architecture. SRC4 lacks both NBS and LRR domains but retains basal antiviral activity through its TIR-SNOR region, with SNOR contributing to TIR-dependent antiviral activity and SRC4 self-association. The EFh domain recognizes coat protein (CP) of soybean mosaic virus (SMV) and serves as a Ca<sup>2+</sup>-responsive module to distinctly regulate TIR-SNOR-mediated immune response in soybean. <i>SRC4-</i>overexpression (Ox-<i>SRC4</i>) soybean plants show SMV resistance without obvious growth penalty. Furthermore, several <i>SRC4</i> high-expression soybean varieties showed stronger SMV resistance and Ca<sup>2+</sup> responsiveness than the <i>SRC4</i> low-expression soybean varieties. Our study highlights the essential roles of atypical resistance proteins in plant immunity and growth-defense balance.</p>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"127 5","pages":""},"PeriodicalIF":6.2,"publicationDate":"2026-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13575729/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148965899","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Yongfang Qin, Congyu Guo, Minseong Kim, Yichuan Zhang, Muhammad Azam, Tingyuan Xiao, Chunhui Xu, Basil Shorrosh, Timothy P. Durrett, Doug K. Allen, Trupti Joshi, Edgar B. Cahoon, Jay J. Thelen, Dong Xu
Plant lipid research depends on accessible databases that connect genes, pathways, and prior literature. However, currently available resources are fragmented, species-limited, and lack AI-powered interfaces for integrated querying. FatPlants 2.0 addresses these issues by integrating ARALIP, PlantFADB, and new Cuphea/Pennycress experimental data into a unified platform with 14 000 genes/proteins, 110 pathways across 5 species, and 57 000+ curated publications. The platform features LipidBot, an AI agent enabling natural language queries via graph-based pathway search and retrieval-augmented generation-powered literature retrieval. Users query complex relationships conversationally and receive answers with traceable citations. The graph database models 100+ biological pathways as queryable networks with large language model-guided Cypher generation. By evaluating more than 1000 curated questions, LipidBot achieved 95% accuracy on pathway queries and 92% recall in literature retrieval using optimized embeddings. The tool demonstrated robust performance across factual, numerical, and multi-hop queries on curated benchmarks. FatPlants 2.0 accelerates research by reducing the time spent on literature reviews. Database and AI agent freely available at https://fatplants.net with bulk downloads and quarterly updates.
{"title":"FatPlants 2.0: an AI-powered platform integrating plant lipid genes, pathways, and literature","authors":"Yongfang Qin, Congyu Guo, Minseong Kim, Yichuan Zhang, Muhammad Azam, Tingyuan Xiao, Chunhui Xu, Basil Shorrosh, Timothy P. Durrett, Doug K. Allen, Trupti Joshi, Edgar B. Cahoon, Jay J. Thelen, Dong Xu","doi":"10.1111/tpj.71117","DOIUrl":"10.1111/tpj.71117","url":null,"abstract":"<p>Plant lipid research depends on accessible databases that connect genes, pathways, and prior literature. However, currently available resources are fragmented, species-limited, and lack AI-powered interfaces for integrated querying. FatPlants 2.0 addresses these issues by integrating ARALIP, PlantFADB, and new Cuphea/Pennycress experimental data into a unified platform with 14 000 genes/proteins, 110 pathways across 5 species, and 57 000+ curated publications. The platform features LipidBot, an AI agent enabling natural language queries via graph-based pathway search and retrieval-augmented generation-powered literature retrieval. Users query complex relationships conversationally and receive answers with traceable citations. The graph database models 100+ biological pathways as queryable networks with large language model-guided Cypher generation. By evaluating more than 1000 curated questions, LipidBot achieved 95% accuracy on pathway queries and 92% recall in literature retrieval using optimized embeddings. The tool demonstrated robust performance across factual, numerical, and multi-hop queries on curated benchmarks. FatPlants 2.0 accelerates research by reducing the time spent on literature reviews. Database and AI agent freely available at https://fatplants.net with bulk downloads and quarterly updates.</p>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"127 5","pages":""},"PeriodicalIF":6.2,"publicationDate":"2026-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13578592/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148975146","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Jing Zhou, Xiaoshuang Zhou, Jiahui Nie, Yetong Qi, Lang Liu, Xinya Wu, Yingtao Zuo, Ruimin Yu, Lei Guo, Paul R.J. Birch, Zhendong Tian
KNOX (KNOTTED1-like HOMEOBOX) transcription factors regulate the expression of genes involved in plant growth, development, and immunity by specifically binding to DNA elements. Previously, we found that the Phytophthora infestans RxLR effector Pi22798 targets the potato transcription factor StKNOX3 to promote colonization. We show here that StKNOX3 interacts with StHUB1 and StHUB2, two E3 ubiquitin ligases that monoubiquitinate histone 2B to form H2Bub1. StKNOX3 stabilizes StHUB1 and StHUB2 in the nucleus, and the StKNOX3-StHUB1/2 complex acts to enhance P. infestans infection. The effector Pi22798 promotes the formation of the StKNOX3-StHUB1/2 complex, facilitating the nuclear accumulation of StHUB1/2, thereby elevating H2Bub1 levels and increasing plant susceptibility. Combined ChIP-Seq and RNA-Seq analyses demonstrate that StKNOX3 modulates transcriptome reprogramming governing cell wall composition, phytohormone responses, and stress responses. StKNOX3 directly binds to the core motifs TGAC or TGTCA in promoter regions of target genes and represses the expression of multiple defense-related genes. Expression of Pi22798 results in reinforced regulation of the StKNOX3-targeted genes by affecting the binding affinity of StKNOX3 to their promoters and increasing H2Bub1 levels. Our data support a model in which the host StKNOX3-StHUB1/2 complex is hijacked by Pi22798 to induce epigenetic modifications that ultimately enhance potato susceptibility to P. infestans.
{"title":"A Phytophthora effector hijacks host StKNOX3-StHUB1/2 complex to reprogram defense gene expression via histone monoubiquitination","authors":"Jing Zhou, Xiaoshuang Zhou, Jiahui Nie, Yetong Qi, Lang Liu, Xinya Wu, Yingtao Zuo, Ruimin Yu, Lei Guo, Paul R.J. Birch, Zhendong Tian","doi":"10.1111/tpj.71112","DOIUrl":"10.1111/tpj.71112","url":null,"abstract":"<p>KNOX (<span>KNO</span>TTED1-like HOMEOBO<span>X</span>) transcription factors regulate the expression of genes involved in plant growth, development, and immunity by specifically binding to DNA elements. Previously, we found that the <i>Phytophthora infestans</i> RxLR effector Pi22798 targets the potato transcription factor StKNOX3 to promote colonization. We show here that StKNOX3 interacts with StHUB1 and StHUB2, two E3 ubiquitin ligases that monoubiquitinate histone 2B to form H2Bub1. StKNOX3 stabilizes StHUB1 and StHUB2 in the nucleus, and the StKNOX3-StHUB1/2 complex acts to enhance <i>P. infestans</i> infection. The effector Pi22798 promotes the formation of the StKNOX3-StHUB1/2 complex, facilitating the nuclear accumulation of StHUB1/2, thereby elevating H2Bub1 levels and increasing plant susceptibility. Combined ChIP-Seq and RNA-Seq analyses demonstrate that StKNOX3 modulates transcriptome reprogramming governing cell wall composition, phytohormone responses, and stress responses. StKNOX3 directly binds to the core motifs TGAC or TGTCA in promoter regions of target genes and represses the expression of multiple defense-related genes. Expression of Pi22798 results in reinforced regulation of the StKNOX3-targeted genes by affecting the binding affinity of StKNOX3 to their promoters and increasing H2Bub1 levels. Our data support a model in which the host StKNOX3-StHUB1/2 complex is hijacked by Pi22798 to induce epigenetic modifications that ultimately enhance potato susceptibility to <i>P. infestans</i>.</p>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"127 5","pages":""},"PeriodicalIF":6.2,"publicationDate":"2026-09-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13575310/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148965845","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Hu Xin, Fang Luo, Dan Zong, Xulei Guo, Richard D. Vierstra, Laigeng Li
Fiber cells are a primary component of vascular tissue in angiosperms, offering mechanical support for most aerial structures, reinforcing vascular transport functions, and enabling vertical growth. Our previous research discovered light as a vital signal in modulating fiber cell-wall thickening and thus length and strength during maturation. Here, we reveal that the red light-sensing Phytochrome B (PhyB)/Phy-Interacting Factor-4 (PIF4) module plays a major photoregulatory role in Arabidopsis stem fiber growth upon activation through its negative influence on the downstream transcription factors encoded by the BR-ENHANCED EXPRESSION (BEE) gene family. Whereas stem fiber cells in the phyB mutant hyperelongate, those in the pif quadruple mutant, which blocks the expression of PIF1-4, are shorter. DNA binding studies showed that this PhyB/PIF module photoregulates BEE2 expression by directly interacting with G-box-like elements within its promoter. Accordingly, fibers in the triple bee123 mutant are shorter, whereas stems overexpressing BEE2 develop longer fibers. BEE2 in particular then encourages fiber cell elongation by directly enhancing expression of genes associated with cell-wall loosening, two being the xyloglucan endotransglycosylases/glucanases XTH8 and XTH33. Our studies identify a key multicomponent photoregulatory cascade in fiber cell elongation, thus offering possible avenues for engineering stem properties by modulating light signals.
{"title":"The PhyB-PIF4 module directly photoregulates BR-ENHANCED EXPRESSION genes to enhance stem fiber cell elongation in Arabidopsis","authors":"Hu Xin, Fang Luo, Dan Zong, Xulei Guo, Richard D. Vierstra, Laigeng Li","doi":"10.1111/tpj.71110","DOIUrl":"10.1111/tpj.71110","url":null,"abstract":"<p>Fiber cells are a primary component of vascular tissue in angiosperms, offering mechanical support for most aerial structures, reinforcing vascular transport functions, and enabling vertical growth. Our previous research discovered light as a vital signal in modulating fiber cell-wall thickening and thus length and strength during maturation. Here, we reveal that the red light-sensing Phytochrome B (PhyB)/Phy-Interacting Factor-4 (PIF4) module plays a major photoregulatory role in Arabidopsis stem fiber growth upon activation through its negative influence on the downstream transcription factors encoded by the <i>BR-ENHANCED EXPRESSION</i> (<i>BEE</i>) gene family. Whereas stem fiber cells in the <i>phyB</i> mutant hyperelongate, those in the <i>pif quadruple</i> mutant, which blocks the expression of <i>PIF1</i>-<i>4</i>, are shorter. DNA binding studies showed that this PhyB/PIF module photoregulates <i>BEE2</i> expression by directly interacting with G-box-like elements within its promoter. Accordingly, fibers in the triple <i>bee123</i> mutant are shorter, whereas stems overexpressing <i>BEE2</i> develop longer fibers. <i>BEE2</i> in particular then encourages fiber cell elongation by directly enhancing expression of genes associated with cell-wall loosening, two being the xyloglucan endotransglycosylases/glucanases XTH8 and XTH33. Our studies identify a key multicomponent photoregulatory cascade in fiber cell elongation, thus offering possible avenues for engineering stem properties by modulating light signals.</p>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"127 5","pages":""},"PeriodicalIF":6.2,"publicationDate":"2026-09-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13575309/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148965867","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Andrew J. Tuckey, Andrew C. Marshall, Emel Rothzerg, Antonella P. Monte, Tom Bennett, Charles S. Bond, Mark T. Waters
Strigolactones are phytohormones that regulate shoot branching and facilitate communication with neighbouring plants, bacteria and arbuscular mycorrhizal fungi. In seed plants, strigolactone perception begins with the enzyme-receptor DWARF14 (D14), an α/β-hydrolase likely to have evolved via gene duplication within a larger gene family that includes the karrikin receptor, KARRIKIN INSENSITIVE 2 (KAI2) and other D14-like (DLK) proteins of uncertain function. However, it is unclear when the functional characteristics that define a bona fide strigolactone receptor evolved. Here, we apply ancestral sequence reconstruction to generate a D14 protein representative of seed plants to study the evolution of ligand specificity and the conservation of signalling mechanisms and to explore desirable traits for protein engineering. Ancestrally reconstructed D14 is structurally and functionally comparable to D14 from Arabidopsis thaliana. The ligand specificity of ancestral D14 does not meaningfully differ, despite having unusual active site geometry and altered enzyme kinetics. We show that the catalytic aspartic acid residue of ancestral D14 is not required for strigolactone signalling when expressed in A. thaliana, suggesting that this mechanistic feature is conserved throughout seed plants. We find that ancestral D14 exhibits higher recombinant yields, greatly increased thermostability and enhanced catalytic activity relative to D14 from A. thaliana. This work provides insight into the evolution of phytohormone signalling and presents a robust scaffold for the application of D14-type proteins in diverse applications.
{"title":"Structural and functional characterisation of a reconstructed ancestral strigolactone receptor","authors":"Andrew J. Tuckey, Andrew C. Marshall, Emel Rothzerg, Antonella P. Monte, Tom Bennett, Charles S. Bond, Mark T. Waters","doi":"10.1111/tpj.71113","DOIUrl":"10.1111/tpj.71113","url":null,"abstract":"<p>Strigolactones are phytohormones that regulate shoot branching and facilitate communication with neighbouring plants, bacteria and arbuscular mycorrhizal fungi. In seed plants, strigolactone perception begins with the enzyme-receptor DWARF14 (D14), an α/β-hydrolase likely to have evolved via gene duplication within a larger gene family that includes the karrikin receptor, KARRIKIN INSENSITIVE 2 (KAI2) and other D14-like (DLK) proteins of uncertain function. However, it is unclear when the functional characteristics that define a <i>bona fide</i> strigolactone receptor evolved. Here, we apply ancestral sequence reconstruction to generate a D14 protein representative of seed plants to study the evolution of ligand specificity and the conservation of signalling mechanisms and to explore desirable traits for protein engineering. Ancestrally reconstructed D14 is structurally and functionally comparable to D14 from <i>Arabidopsis thaliana</i>. The ligand specificity of ancestral D14 does not meaningfully differ, despite having unusual active site geometry and altered enzyme kinetics. We show that the catalytic aspartic acid residue of ancestral D14 is not required for strigolactone signalling when expressed in <i>A. thaliana</i>, suggesting that this mechanistic feature is conserved throughout seed plants. We find that ancestral D14 exhibits higher recombinant yields, greatly increased thermostability and enhanced catalytic activity relative to D14 from <i>A. thaliana</i>. This work provides insight into the evolution of phytohormone signalling and presents a robust scaffold for the application of D14-type proteins in diverse applications.</p>","PeriodicalId":233,"journal":{"name":"The Plant Journal","volume":"127 5","pages":""},"PeriodicalIF":6.2,"publicationDate":"2026-09-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13575467/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148965855","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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