Plant Science最新文献_第4页

Transcription factor PdMYB118 in poplar regulates lignin deposition and xylem differentiation in addition to anthocyanin synthesis through suppressing the expression of PagKNAT2/6b gene 杨树中的转录因子 PdMYB118 除了通过抑制 PagKNAT2/6b 基因的表达来调节木质素沉积和木质部分化外，还能调节花青素的合成。

IF 4.2 2区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Plant Science

Pub Date : 2024-10-09 DOI: 10.1016/j.plantsci.2024.112277

Shuo Song , Wei Guo , Yu Guo , Erkun Chao , Sujie Sun , Lizi Zhao , Yanqiu Zhao , Hongxia Zhang

R2R3-MYB transcription factors function as the master regulators of the phenylpropanoid pathway in which both lignin and anthocyanin are produced. In poplar, R2R3-MYB transcription factor PdMYB118 positively regulates anthocyanin production to change leaf color. However, the molecular mechanism by which it controls different branches of the phenylpropanoid pathway still remains poorly understood. Here, we reported that in addition to anthocyanin synthesis, lignin deposition and xylem differentiation were regulated by PdMYB118 through inhibiting PagKNAT2/6b gene expression. The transgenic poplar plants overexpressing PdMYB118 accumulated more xylem, lignin and anthocyanin. Transcriptome and reverse transcription quantitative PCR analyses revealed that the expression of PagKNAT2/6b gene which inhibited lignin deposition and xylem differentiation was significantly down-regulated in transgenic poplar plants. Subsequent dual-luciferase reporter and yeast-one-hybrid assays demonstrated that PdMYB118 directly inhibited the transcription of PagKNAT2/6b by binding to the AC elements in its promoter region. Further experiments with transgenic poplar plants overexpressing PagKNAT2/6b demonstrated that overexpression of PagKNAT2/6b in the PdMYB118 overexpression background rescued lignin accumulation and xylem width to the same level of wild type plants. The findings in this work suggest that PdMYB118 is involved in the lignin deposition and xylem differentiation via modulating the expression of PagKNAT2/6b, and the PdMYB118- PagKNAT2/6b model can be used for the genetic breeding of new woody tree with high lignin production.

R2R3-MYB 转录因子是产生木质素和花青素的苯丙酮途径的主调节因子。在杨树中，R2R3-MYB 转录因子 PdMYB118 能正向调节花青素的产生，从而改变叶片颜色。然而，人们对其控制苯丙酮途径不同分支的分子机制仍然知之甚少。在此，我们报道了 PdMYB118 除了调控花青素的合成外，还通过抑制 PagKNAT2/6b 基因的表达来调控木质素的沉积和木质部的分化。过表达 PdMYB118 的转基因杨树植株积累了更多的木质部、木质素和花青素。转录组和逆转录定量 PCR 分析表明，在转基因杨树植株中，抑制木质素沉积和木质部分化的 PagKNAT2/6b 基因表达显著下调。随后进行的双荧光素酶报告和酵母一杂交试验表明，PdMYB118 通过与 PagKNAT2/6b 启动子区域中的 AC 元件结合，直接抑制了 PagKNAT2/6b 的转录。用过表达 PagKNAT2/6b 的转基因杨树植株进行的进一步实验表明，在 PdMYB118 过表达背景下，过表达 PagKNAT2/6b 可将木质素积累和木质部宽度恢复到与野生型植株相同的水平。该研究结果表明，PdMYB118通过调节PagKNAT2/6b的表达参与木质素沉积和木质部分化，PdMYB118- PagKNAT2/6b模型可用于遗传育种高木质素产量的新型木本植物。

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引用次数: 0

VvRF2b interacts with VvTOR and influences VvTOR-regulated sugar metabolism in grape VvRF2b 与 VvTOR 相互作用并影响 VvTOR 调控的葡萄糖代谢。

IF 4.2 2区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Plant Science

Pub Date : 2024-10-01 DOI: 10.1016/j.plantsci.2024.112276

Shuang Xia , Ying Zhao , Qiaoyun Deng , Xiaoyu Han , Xiuqin Wang

The production of top-quality wines is closely related to the quality of the wine grapes. In wine grapes (Vitis vinifera L., Vv), sugar is a crucial determinant of berry quality, regulated by an interplay of various transcription factors and key kinases. Many transcription factors involved in sugar metabolism remain unexplored. Target of Rapamycin (TOR) is an important protein kinase in plants, recently found to regulate sugar metabolism in grapes. However, transcription factors or other factors involved in this process are rarely reported. Here, we utilized transgenic callus tissues from 'Cabernet Sauvignon' grape fruit engineered via gene overexpression (oe) and CRISPR/Cas9-based gene knockout (ko), and discovered a bZIP transcription factor, VvRF2b, whose knockout resulted in increased accumulation of fructose and sucrose, indicating that VvRF2b is a negative regulator of sugar accumulation. Subcellular localization and transcriptional activation tests showed that VvRF2b is an activator of transcription located both in the nucleus and cell membrane. Analysis of VvRF2b and VvTOR gene levels and sugar contents (glucose, fructose, and sucrose) in 'Cabernet Sauvignon' grape fruits at 30, 70, and 90 days after bloom (DAB) revealed that VvRF2b is expressed more highly during fruit development, while VvTOR is expressed more during the sugar accumulation phase, furthermore, VvTOR gene levels in koVvRF2b transgenic calli increased significantly, suggesting a strong relationship between the knockout of VvRF2b and the overexpression of VvTOR. Additionally, bimolecular fluorescence complementation and luciferase complementation assays demonstrated the interaction between VvRF2b and VvTOR proteins. After knocking out the VvRF2b gene in oeVvTOR calli, it was found that the knockout of VvRF2b promotes VvTOR-regulated sucrose accumulation and enhances the expression of sugar metabolism-related genes regulated by VvTOR. In summary, our results suggest that VvRF2b interacts with VvTOR protein and influences VvTOR-regulated sugar metabolism.

顶级葡萄酒的生产与酿酒葡萄的质量密切相关。在酿酒葡萄（Vitis vinifera L., Vv）中，糖分是决定浆果质量的关键因素，受各种转录因子和关键激酶的相互作用调节。许多参与糖代谢的转录因子仍未得到研究。雷帕霉素靶蛋白激酶（TOR）是植物中一种重要的蛋白激酶，最近发现它能调节葡萄的糖代谢。然而，参与这一过程的转录因子或其他因子却鲜有报道。在这里，我们利用'赤霞珠'葡萄果实的转基因胼胝体组织，通过基因过表达（oe）和基于CRISPR/Cas9的基因敲除（ko），发现了一种bZIP转录因子VvRF2b，其敲除会导致果糖和蔗糖的积累增加，表明VvRF2b是糖积累的负调控因子。亚细胞定位和转录激活测试表明，VvRF2b 是位于细胞核和细胞膜的转录激活因子。对开花后 30 天、70 天和 90 天（DAB）'赤霞珠'葡萄果实中 VvRF2b 和 VvTOR 基因水平和糖分含量（葡萄糖、果糖和蔗糖）的分析表明，VvRF2b 在果实发育过程中表达较多、此外，在 koVvRF2b 转基因胼胝体中，VvTOR 基因水平显著增加，这表明 VvRF2b 基因敲除与 VvTOR 基因过表达之间存在密切关系。此外，双分子荧光互补和荧光素酶互补实验证明了 VvRF2b 和 VvTOR 蛋白之间的相互作用。在 oeVvTOR 胼胝体中敲除 VvRF2b 基因后发现，敲除 VvRF2b 能促进 VvTOR 调控的蔗糖积累，并增强 VvTOR 调控的糖代谢相关基因的表达。总之，我们的研究结果表明，VvRF2b 与 VvTOR 蛋白相互作用，并影响 VvTOR 调控的糖代谢。

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引用次数: 0

Application of uniconazole in improving the high-throughput genetic transformation efficiency in maize 在提高玉米高通量遗传转化效率中应用联苯苄唑。

IF 4.2 2区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Plant Science

Pub Date : 2024-09-28 DOI: 10.1016/j.plantsci.2024.112270

Shengnan Liu , Jihui Qiao , Shuaisong Zhang , Minhui Lu , Yongqing Yang , Jinsheng Lai , Yan Guo , Yunlu Shi

Agrobacterium-mediated genetic transformation is the most effective and widely used delivery system for candidate genes and genome editors in maize, which is an important crop with the largest planting area and the highest yield. Here, we used gibberellin synthesis inhibitor, uniconazole, to enhance the stem strength of regenerated plantlets resulting in a significantly increase from 11.6 % to 18.2 % in the percentage of regenerated plantlets, and the transformation frequency was also improved from 9.4 % to 15.6 % in the test experiments. The physiological condition of immature embryo is greatly affected by ear source, season and insect pests, while it can cause significant fluctuations in the transformation frequency. Our optimization works at the differentiation subculture stage, avoiding the impact on the physiological condition of immature embryo. So, it can be applicated to high-throughput genetic transformation in different seasons and different ear sources throughout the year. The productive experiment results indicated that the annual average transformation frequency significantly improved from 2.76 % to 7.14 % (approximately 2.6 folds improvement), and the tissue culture cycle was shortened from 115 days to 106 days by using optimized system. Our optimized genetic transformation system opens avenues for maize improvement based on transgenic and genome editing technology.

玉米是种植面积最大、产量最高的重要作物，农杆菌介导的遗传转化是玉米候选基因和基因组编辑最有效、应用最广泛的传递系统。在这里，我们使用赤霉素合成抑制剂--烯效唑来增强再生小苗的茎干强度，结果再生小苗的比例从 11.6% 显著提高到 18.2%，转化频率也从 9.4% 提高到 15.6%。未成熟胚的生理状况受穗源、季节和虫害的影响很大，同时也会导致转化频率的大幅波动。我们的优化方案在分化亚培养阶段就能发挥作用，避免了对未成熟胚胎生理状况的影响。因此，它可以应用于全年不同季节和不同耳源的高通量遗传转化。生产性实验结果表明，使用优化系统后，年平均转化率从 2.76% 显著提高到 7.14%（约提高 2.6 倍），组织培养周期从 115 天缩短到 106 天。我们的优化遗传转化系统为基于转基因和基因组编辑技术的玉米改良开辟了道路。

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引用次数: 0

Beneficial or detrimental? How nickel application alters the ionome of soybean plants 有益还是有害？施用镍如何改变大豆植物的离子组？

IF 4.2 2区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Plant Science

Pub Date : 2024-09-27 DOI: 10.1016/j.plantsci.2024.112274

Daniel Venâncio de Paula Correia , Bruna Wurr Rodak , Henrique Amorim Machado , Guilherme Lopes , Douglas Siqueira Freitas

The use of nickel (Ni) in agriculture may represent one of the most significant cases of plant hormesis ever reported, as plants exhibit both positive and negative responses depending on the level of exposure to this element. For a more comprehensive understanding of this effect, the next step is to conduct studies on the dynamics of pre-existing chemical elements in the system (ionomic profile), especially when introducing Ni as a novel nutrient for the plants. This micronutrient is of particular interest to the fertilization of leguminous plants, such as the soybean, due to its additional effects on the biological nitrogen fixation process. This study thus evaluated the influence of five doses of Ni (0.0, 0.5, 1.0, 3.0, and 9.0 mg of Ni kg⁻¹) on the ionomic profile of soybean genotypes using modern quantification techniques. The results revealed that the addition of Ni reduced the concentration of cationic micronutrients manganese (Mn), iron (Fe), zinc (Zn), and copper (Cu), while it increased the concentration of macronutrients nitrogen (N) and magnesium (Mg). The application of Ni also resulted in a reduction of the potentially toxic element aluminum (Al). Correlations were also observed for these elements, indicating that Ni could be a controlling agent in elemental absorption and translocation. The ionome of the leaf tissues exhibited the most significant alterations, followed by the grains, nodules, and roots. Exogenous agronomic doses of Ni proved beneficial for the growth and production of soybean plants, although a genotypic effect was observed. The treatment with 9.0 mg of Ni kg⁻¹, resulted in a new ionomic profile related to toxicity, demonstrating suboptimal plant development. Thus, the application of Ni in appropriate doses had a significant impact on the ionomic profile of soybeans, improving plant development and implying resistance to potentially toxic elements such as Al.

镍（Ni）在农业中的应用可能是有报道以来最重要的植物激素作用案例之一，因为植物会根据接触这种元素的程度表现出积极和消极的反应。为了更全面地了解这种效应，下一步是对系统中原有化学元素的动态（离子组态）进行研究，尤其是在引入镍作为植物的新营养元素时。这种微量营养元素对豆科植物（如大豆）的施肥尤为重要，因为它对生物固氮过程有额外的影响。因此，本研究利用现代定量技术评估了五种剂量的镍（0.0、0.5、1.0、3.0 和 9.0 毫克镍 kg-1）对大豆基因型离子组学特征的影响。结果表明，添加镍降低了阳离子微量营养元素锰（Mn）、铁（Fe）、锌（Zn）和铜（Cu）的浓度，而增加了大量营养元素氮（N）和镁（Mg）的浓度。施用镍还能减少潜在有毒元素铝（Al）的含量。还观察到这些元素之间的相关性，表明镍可能是元素吸收和转移的控制剂。叶片组织的离子组发生了最显著的变化，其次是谷粒、瘤和根。事实证明，外源农艺剂量的镍有利于大豆植株的生长和产量，但也存在基因型效应。9.0mg Ni kg-1 的处理导致了与毒性有关的新的离子组学特征，显示出植物发育不理想。因此，施用适当剂量的镍对大豆的植株特征有显著影响，能改善植株发育，并对铝等潜在有毒元素产生抗性。

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引用次数: 0

AhGSNOR1 negatively regulates Al-induced programmed cell death by regulating intracellular NO and redox levels AhGSNOR1 通过调节细胞内 NO 和氧化还原水平，对 Al 诱导的细胞程序性死亡进行负向调节。

IF 4.2 2区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Plant Science

Pub Date : 2024-09-26 DOI: 10.1016/j.plantsci.2024.112275

Chunliu Pan , Xia Li , Changge Jian , Yunyi Zhou , Aiqin Wang , Dong Xiao , Jie Zhan , Longfei He

The toxicity of aluminum (Al) in acidic soil inhibits plant development and reduces crop yields. Programmed cell death (PCD) is one of the important mechanisms in the plant response to Al toxicity. However, it is yet unknown if S-nitrosoglutathione reductase (GSNOR) provides Al-PCD. Here, transcription and protein expression of AhGSNOR1 were both induced by Al stress. AhGSNOR1-overexpressing transgenic tobacco plants reduced Al-induced nitric oxide (NO) and S-nitrosothiol accumulation, the inhibitory effect of Al stress on root elongation and the degree of cell death, and enhanced antioxidant enzyme activity to effectively remove hydrogen peroxide. In addition, AhGSNOR1 directly interacted with AhTRXh in vivo. Expression of Trxh3 in AhGSNOR1-overexpressing transgenic plants was significantly upregulated, indicating that AhGSNOR1 positively regulated the transcriptional level of Trxh3. Together, these results suggested that AhGSNOR1 was a negative regulatory factor of Al-induced PCD and improved plant Al-tolerance by modulating intracellular NO and redox homeostasis.

酸性土壤中的铝（Al）毒性会抑制植物生长并降低作物产量。程序性细胞死亡（PCD）是植物应对铝毒性的重要机制之一。然而，S-亚硝基谷胱甘肽还原酶（GSNOR）是否能提供铝-程序性细胞死亡尚不清楚。在这里，AhGSNOR1的转录和蛋白表达都受到了Al胁迫的诱导。AhGSNOR1-overexpressing 转基因烟草植株减少了铝诱导的一氧化氮（NO）和 S-亚硝基硫醇的积累，降低了铝胁迫对根系伸长的抑制作用和细胞死亡程度，并增强了抗氧化酶的活性，从而有效清除过氧化氢。此外，AhGSNOR1在体内直接与AhTRXh相互作用。在 AhGSNOR1 基因缺失的转基因植株中，Trxh3 的表达显著上调，表明 AhGSNOR1 能正向调节 Trxh3 的转录水平。这些结果表明，AhGSNOR1是铝诱导的PCD的负调控因子，并通过调节细胞内NO和氧化还原平衡提高植物对铝的耐受性。

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引用次数: 0

Analysis of the role of the rice metallothionein gene OsMT2b in grain size regulation 水稻金属硫蛋白基因 OsMT2b 在粒度调节中的作用分析

IF 4.2 2区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Plant Science

Pub Date : 2024-09-24 DOI: 10.1016/j.plantsci.2024.112272

Jian Chen, Yunyi Wen, Yibin Pan, Ying He, Xiaoting Gong, Wenli Yang, Weiting Chen, Feng Zhou, Dagang Jiang

Seed size is one of the three main characteristics determining rice yield. Clarification of the mechanisms regulating seed size in rice has implications for improving rice yield. Although several genes have been reported to regulate seed size, most of the reports are fragmentary. The role of metallothioneins (MTs) in regulating seed size remains unknown. Here, we found that OsMT2b was expressed in both spikelets and developing seeds. OsMT2b-overexpression lines had large and heavy seeds, and RNAi (RNA interference) lines had small and light seeds. Scanning electron microscopy (SEM) observations revealed that OsMT2b regulated spikelet hull size by affecting cell expansion in the outer epidermis. Histological analysis indicated that OsMT2b affected the number of cells in the cross-section of spikelet hulls, which affected seed size. The fresh weight of seeds was consistently higher in OsMT2b-overexpression lines than in seeds of the wild-type (WT) and RNAi lines from 6 DAP (days after pollination) until maturity, indicating that OsMT2b affected seed filling. Reverse transcription-quantitative PCR (RT-qPCR) analyses revealed that OsMT2b regulates the expression of reactive oxygen species scavenging-related genes involved in seed size regulation. In conclusion, our results indicated that OsMT2b positively regulates seed size, which provides a novel approach for regulating seed size with genetic engineering technology.

种子大小是决定水稻产量的三大特征之一。阐明水稻种子大小的调控机制对提高水稻产量具有重要意义。虽然已有多个基因被报道调控种子大小，但大多数报道都很零散。金属硫蛋白（MTs）在调节种子大小中的作用仍然未知。在这里，我们发现 OsMT2b 在小穗和发育中的种子中都有表达。OsMT2b 高表达株系的种子大而重，而 RNAi（RNA 干扰）株系的种子小而轻。扫描电子显微镜（SEM）观察发现，OsMT2b 通过影响外表皮细胞的扩张来调节小穗外壳的大小。组织学分析表明，OsMT2b 会影响小穗外壳横截面上的细胞数量，从而影响种子大小。从 6 DAP（授粉后天数）到成熟，OsMT2b 高表达株系的种子鲜重一直高于野生型（WT）和 RNAi 株系的种子，这表明 OsMT2b 影响了种子的充实。反转录定量 PCR（RT-qPCR）分析表明，OsMT2b 可调控参与种子大小调控的活性氧清除相关基因的表达。总之，我们的研究结果表明，OsMT2b 能正向调节种子大小，这为利用基因工程技术调节种子大小提供了一种新方法。

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引用次数: 0

An osmesl mutant delayed rice leaf senescence through inhibiting cell death by OsBI-1 osmesl突变体通过抑制OsBI-1导致的细胞死亡来延缓水稻叶片衰老。

IF 4.2 2区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Plant Science

Pub Date : 2024-09-23 DOI: 10.1016/j.plantsci.2024.112271

Bin Hu , Enlong Shen , Fengling Zhou , Bo Sun , Xingchao Wang , Fei Zhou , Yongjun Lin

Leaf senescence following heading in rice is subject to rigorous regulation, with many of the underlying control mechanisms remaining largely unknown. In this study, we identified a novel gene, OsMESL, which exerts a positive regulatory effect on leaf senescence in rice. The T-DNA insertion mutant known as osmesl and RNA interference plants displayed a phenotype characterized by stay-green after heading. Genetic analysis indicated that the mutant phenotype could be rescued through complementation, while the overexpression of OsMESL accelerated leaf senescence after heading, underscoring OsMESL's positive regulatory role in rice leaf senescence. Subsequent investigations revealed that OsMESL modulates the process of cell death by influencing the stability of its interacting protein, the cell death suppressor OsBI-1, thereby governing leaf senescence. Furthermore, the leaves of the osmesl mutant exhibited a delayed reduction in photosynthesis, along with increased grain length and 1000-grain weight. In conclusion, we identified OsMESL as a novel positive regulator of leaf senescence in rice, which likely participates in leaf senescence through the mediation of cell death by OsBI-1, resulting in the phenotype of stay-green in the osmesl mutant after heading.

水稻萌发后的叶片衰老受到严格的调控，许多潜在的调控机制在很大程度上仍然未知。在这项研究中，我们发现了一个新基因 OsMESL，它对水稻的叶片衰老具有积极的调控作用。T-DNA插入突变体osmesl和RNA干扰植株表现出的表型特征是头后留绿。遗传分析表明，该突变体的表型可通过互补得到挽救，而 OsMESL 的过表达则会加速打顶后的叶片衰老，这突显了 OsMESL 在水稻叶片衰老中的正向调控作用。随后的研究发现，OsMESL通过影响与其相互作用的蛋白--细胞死亡抑制因子OsBI-1的稳定性来调节细胞死亡过程，从而控制叶片衰老。此外，Osmesl突变体的叶片表现出光合作用延迟降低，同时谷粒长度和千粒重增加。总之，我们发现 OsMESL 是一种新型的水稻叶片衰老正调控因子，它可能通过 OsBI-1 对细胞死亡的调控参与叶片衰老，从而导致 osmesl 突变体在打顶后出现留绿表型。

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引用次数: 0

A vacuolar protein MaSCPL1 mediates anthocyanin acylation modifications in blue-flowered grape hyacinth 一种液泡蛋白 MaSCPL1 在蓝花葡萄风信子中介导花青素酰化修饰

IF 4.2 2区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Plant Science

Pub Date : 2024-09-23 DOI: 10.1016/j.plantsci.2024.112273

Xiaoyun Cao , Wenhui Hao , Wanqi Pan , Xuelan Gao , Jingwen Xie , Lingjuan Du

The grape hyacinth is renowned for its profuse blue flowers, which confer substantial scientific and ornamental significance as well as considerable potential for industrial applications. The serine carboxypeptidase-like acyltransferases (SCPL-ATs) family is crucial for the blue flower coloration. To elucidate SCPL-ATs involved in anthocyanin modification in grape hyacinth, we performed a transcriptomic analysis of grape hyacinth SCPL-ATs. Through gene expression profiling, we identified a promising candidate gene, MaSCPL1, whose expression patterns corresponded with variations in anthocyanin content throughout petal coloration. Subsequently, the functional role of the MaSCPL1 gene was validated using the native petal regeneration system, and the silencing of MaSCPL1 led to a decreased total anthocyanin content and Dp3MG content in grape hyacinth petals. Furthermore, we employed yeast one-hybrid (Y1H), electrophoretic mobility shift assay (EMSA), and dual-luciferase assays to explore the regulatory interactions between the anthocyanin biosynthesis transcription factor MaMybA and the MaSCPL1 promoter. Our findings indicate that MaMybA can bind to the MaSCPL1 promoter and significantly activate its expression. Furthermore, the MaMybA-RNAi resulted in a substantial multifold reduction in the expression of MaSCPL1, implying that the regulation of MaSCPL1 expression is mediated by MaMybA. This study revealed the MaSCPL1 gene has been associated with anthocyanin acylated modification in grape hyacinth and elucidated the important role of the MaMybA-MaSCPL1 module in colouration grape hyacinth.

葡萄风信子因其盛开的蓝色花朵而闻名于世，这种花朵不仅具有重要的科学和观赏价值，而且在工业应用方面也具有相当大的潜力。丝氨酸羧肽酶样酰基转移酶（SCPL-ATs）家族对蓝色花朵的着色至关重要。为了阐明参与葡萄风信子花青素修饰的 SCPL-ATs，我们对葡萄风信子 SCPL-ATs 进行了转录组分析。通过基因表达谱分析，我们发现了一个很有希望的候选基因 MaSCPL1，其表达模式与整个花瓣着色过程中花青素含量的变化相对应。随后，我们利用原生花瓣再生系统验证了 MaSCPL1 基因的功能作用，沉默 MaSCPL1 基因会导致葡萄风信子花瓣中总花青素含量和 Dp3MG 含量的降低。此外，我们还采用了酵母单杂交（Y1H）、电泳迁移实验（EMSA）和双荧光素酶实验来探讨花青素生物合成转录因子MaMybA与MaSCPL1启动子之间的调控相互作用。我们的研究结果表明，MaMybA 能与 MaSCPL1 启动子结合并显著激活其表达。此外，MaMybA-RNAi导致MaSCPL1的表达量大幅降低数倍，这意味着MaSCPL1的表达调控是由MaMybA介导的。该研究揭示了MaSCPL1基因与葡萄风信子花青素酰化修饰有关，并阐明了MaMybA-MaSCPL1模块在葡萄风信子着色过程中的重要作用。

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引用次数: 0

Activity and gene expression analysis of the NADP-dependent isocitrate dehydrogenase (NADP-ICDH) through pepper fruit ripening and its modulation by nitric oxide (NO). Molecular characterization of the peroxisomal isozyme 辣椒果实成熟过程中依赖 NADP 的异柠檬酸脱氢酶（NADP-ICDH）的活性和基因表达分析及其受一氧化氮（NO）的调节。过氧化物酶同工酶的分子特征。

IF 4.2 2区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Plant Science

Pub Date : 2024-09-21 DOI: 10.1016/j.plantsci.2024.112269

María A. Muñoz-Vargas, Salvador González-Gordo, Jorge Taboada, José M. Palma, Francisco J. Corpas

NADP-dependent isocitrate dehydrogenase (NADP-ICDH) is one of the main sources of cellular reductant capacity in the form of NADPH. Although there is significant knowledge about the relevance of this enzyme during some physiological and stress processes, the available information about its involvement in fruit ripening is scarce. Using sweet green pepper (Capsicum annuum L.) fruits, a 50–75 % ammonium-sulfate-enriched protein fraction containing the NADP-ICDH activity allowed its biochemical characterization. The enzyme displayed a typical Michaelis-Menten kinetics and exhibited V_max and K_m values of 97 μUnits and 78 µM for isocitrate, and 92 μUnits and 46 µM for NADP⁺. Three NADP-ICDH isozymes were identified by non-denaturing PAGE designated as NADP-ICDH I to III, each representing 33 %, 24 %, and 43 %, respectively, of the total activity. Based on our previous transcriptome (RNA-Seq), three CaICDH genes (CaNADP-ICDH1, CaNADP-ICDH2, and CaNADP-ICDH3) were identified in sweet pepper fruits encoding isozymes potentially distributed in the cytosol, cytosol/mitochondrion, and peroxisome, according to their percentage of identity with the Arabidopsis isozymes. The time-course expression analysis of these genes during different fruit ripening stages including green immature (G), breaking point (BP), and red ripe (R), and in fruits subjected to nitric oxide (NO) treatments, showed dissimilar expression patterns. During ripening from green to red fruits, CaNADP-ICDH1 and CaNADP-ICDH2 were upregulated but were negatively affected by NO; however, CaNADP-ICDH3 was downregulated during ripening but unaffected by NO treatment. Furthermore, during ripening, the NADP-ICDH activity increased in red ripe fruits whereas the NO gas treatment produced a significant inhibition. These findings provide, to our knowledge, the first characterization of the NADP-ICDH family in this non-climacteric fruit and suggest that NADP-ICDH must play an important role in maintaining the supply of NADPH during pepper fruit ripening and that NO partially modulates this NADPH-generating system.

依赖 NADP 的异柠檬酸脱氢酶（NADP-ICDH）是以 NADPH 形式存在的细胞还原剂能力的主要来源之一。尽管人们对这种酶在某些生理和应激过程中的相关性有大量了解，但有关它参与果实成熟的现有信息却很少。利用甜青椒（Capsicum annuum L.）果实，对含有 NADP-ICDH 活性的 50-75% 富含硫酸铵的蛋白质部分进行了生化鉴定。该酶表现出典型的 Michaelis-Menten 动力学，异柠檬酸的 Vmax 和 Km 值分别为 97 μUits 和 78µM，NADP+ 的 Vmax 和 Km 值分别为 92 μUits 和 46µM。通过非变性 PAGE 鉴定出三种 NADP-ICDH 同工酶，命名为 NADP-ICDH I 至 III，分别占总活性的 33%、24% 和 43%。根据我们之前的转录组（RNA-Seq），在甜椒果实中发现了三个 CaICDH 基因（CaNADP-ICDH1、CaNADP-ICDH2 和 CaNADP-ICDH3），根据它们与拟南芥同工酶的相同百分比，这些基因编码的同工酶可能分布在细胞质、细胞质/半胱体和过氧物酶体中。对这些基因在不同果实成熟阶段（包括绿色未成熟果实（G）、破裂点果实（BP）和红色成熟果实（R））以及一氧化氮（NO）处理的果实中的时程表达分析显示出不同的表达模式。在从青果到红果的成熟过程中，CaNADP-ICDH1 和 CaNADP-ICDH2 表达上调，但受到 NO 的负面影响；然而，CaNADP-ICDH3 在成熟过程中表达下调，但不受 NO 处理的影响。此外，在成熟过程中，红熟果实中的 NADP-ICDH 活性增加，而 NO 气体处理会产生明显的抑制作用。据我们所知，这些发现首次描述了这种非成熟果实中 NADP-ICDH 家族的特征，并表明在辣椒果实成熟过程中，NADP-ICDH 必须在维持 NADPH 供应方面发挥重要作用，而 NO 可部分调节这一 NADPH 生成系统。

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引用次数: 0

BjuA03.BNT1 plays a positive role in resistance to clubroot disease in resynthesized Brassica juncea L. BjuA03.BNT1在再合成芸薹属植物抗球根病中发挥积极作用

IF 4.2 2区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Plant Science

Pub Date : 2024-09-21 DOI: 10.1016/j.plantsci.2024.112268

Keqi Li , Kai Wang , Yiji Shi , Fenghao Liang , Xinru Li , Shunjun Bao , Balziya Maratkyzy Yesmagul , Maliha Fatima , Chengyu Yu , Aixia Xu , Xingguo Zhang , Sanxiong Fu , Xue Shi , Xiaoling Dun , Zhaoyong Zhou , Zhen Huang

Clubroot has become a major obstacle in rapeseed production. Breeding varieties resistant to clubroot is the most effective method for disease management. However, the clubroot-resistant germplasm of rapeseed remains limited. To tackle this challenge, we synthesized the clubroot-resistant mustard, CT19, via distant hybridization, and subsequently an F₂ segregating population was created by intercrossing CT19 with a clubroot-susceptible germplasm CS15. A major-effect clubroot resistance QTL qCRa3–1 on chromosome A03 was identified through QTL scanning. Transcriptome analyses of CT19 and CS15 revealed that the mechanisms conferring resistance to Plasmodiophora brassica likely involved the regulation of flavonoid metabolism, fatty acid metabolism, and sulfur metabolism. By combining the results from transcriptome, QTL mapping, and gene sequencing, a candidate gene BjuA03.BNT1, encoding NLR (nucleotide-binding domain leucine-rich repeat-containing receptors) protein, was obtained. Intriguingly, comparing with CT19, a base T insertion was discovered in the BjuA03.BNT1 gene's coding sequence in CS15, resulting an alteration within the LRR conserved domain. Overexpression of BjuA03.BNT1 from CT19 notably enhanced the resistance to clubroot in Arabidopsis. Our investigations revealed that BjuA03.BNT1 regulated the resistance to clubroot by modulating fatty acid synthesis and the structure of cell wall. These results are highly relevant for molecular breeding to improve clubroot resistance in rapeseed.

棒根病已成为油菜籽生产的一大障碍。培育抗倒伏品种是防治病害的最有效方法。然而，抗倒伏油菜种质资源仍然有限。为了应对这一挑战，我们通过远缘杂交合成了抗棒子病的芥菜品种 CT19，然后通过 CT19 与棒子病易感种质 CS15 杂交产生了 F2 分离群体。通过 QTL 扫描，在染色体 A03 上发现了一个主要效应的抗棒根病 QTL qCRa3-1。对CT19和CS15的转录组分析表明，赋予黄花菜抗性的机制可能涉及黄酮代谢、脂肪酸代谢和硫代谢的调控。结合转录组、QTL图谱和基因测序的结果，得到了编码NLR（核苷酸结合域富含亮氨酸重复受体）蛋白的候选基因BjuA03.BNT1。耐人寻味的是，与 CT19 相比，CS15 的 BjuA03.BNT1 基因编码序列中发现了一个碱基 T 插入，导致 LRR 保守结构域发生改变。过表达 CT19 中的 BjuA03.BNT1 能显著增强拟南芥对棒根病的抗性。我们的研究发现，BjuA03.BNT1 通过调节脂肪酸的合成和细胞壁的结构来调控拟南芥对棍棒病的抗性。这些结果对分子育种提高油菜抗倒伏能力具有重要意义。

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引用次数: 0