Plant Signaling & Behavior
·
2022
·
IF 2.734
Rui Zhou,Menglong Fan,Mei Zhao,Xinqiang Jiang,Qinghua Liu
Leaves are the main vegetative organs of the aboveground part of plants and play an important role in plant morphogenesis. KNOTTED-LIKE HOMEOBOX (KNOX) plays a crucial role in regulating leaf cell fate and maintaining leaf development. In this study, we analyzed LtKNOX1 from Lilium tsingtauense and illustrated its function in transgenic plants. Tissue-specific expression analysis indicated that LtKNOX1 was highly expressed in stems, young flower buds, and shoot apical meristems (SAMs). Ectopic overexpression of LtKNOX1 in Nicotiana benthamiana suggested that transformants with mild phenotypes were characterized by foliar wrinkles and mildly curled leaves; transformants with intermediate phenotypes showed severely crimped blades and narrow leaf angles, and the most severe phenotypes lacked normal SAMs and leaves. Moreover, the expression levels of genes involved in the regulation of KNOX in transgenic plants were detected, including ASYMMETRIC LEAVES1, PIN-FORMED 1, GA20-oxidase, CUP-SHAPED COTYLEDON 2, CLAVATA 1 and WUSCHEL(WUS), and the expression of other genes were down-regulated except WUS. This study contributes to our understanding of the LtKNOX1 function.
研究领域:
植物学
·
遗传学
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DOI
PubMed
高影响因子
PEST MANAGEMENT SCIENCE
·
2024
·
IF 3.8
Xin-Yue Zhang,Wen-Hao Han,Feng-Bin Zhang,Jun-Xia Wang,Shu-Sheng Liu,Xiao-Wei Wang
BACKGROUND The whitefly Bemisia tabaci is one of the world's foremost agricultural pests. Recently, we found that a wild relative of tobacco ( Nicotiana benthamiana ) demonstrates remarkable attractiveness and nearly 100% lethality towards whiteflies. Therefore, it can act as a dead-end trap crop for whitefly control in the field. However, the underlying mechanism of the significant attractiveness of N. benthamiana towards whiteflies is unclear. RESULTS Binary-choice assays and olfactory experiments showed that compared to common tobacco ( N. tabacum ), the volatile of N. benthamiana has a greater attraction to whiteflies. Then we analyzed and compared volatiles from these two Nicotiana species by thermal desorption-gas chromatography–mass spectrometry (TD-GC–MS). We identified 16 chemical compounds that are more abundant in N. benthamiana than in N. tabacum. Seven compounds were further tested with olfactometer assays and we found that, among them, undecane strongly attracted whiteflies. Further experiments revealed that even 0.005 μg mL −1 undecane is attractive to whiteflies. We also silenced the genes that may influence the biosynthesis of undecane and found the production of undecane decreased after silencing NbCER3 , and that N. benthamiana plants with less undecane lost their attraction to whiteflies. In addition, we found that applying 0.005 μg mL −1 undecane on yellow sticky traps can increase the number of stuck insects on the traps by ≈40%. CONCLUSION Undecane from the volatile of N. benthamiana is a critical chemical signal that attracts whiteflies and NbCER3 involved in the biosynthesis of undecane. Undecane may be used to improve the efficiency of yellow sticky traps for whitefly control. © 2024 Society of Chemical Industry.
研究领域:
农业昆虫学
·
植物科学
·
化学生态学
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DOI
PubMed
高影响因子
MOLECULAR PLANT PATHOLOGY
·
2021
·Vol. 23
·期 1
·60-77
·
IF 5.663
Changyun Liu,Shaorui Tian,Xing Lv,Yundan Pu,Haoran Peng,Guangjin Fan,Xiaozhou Ma,Lisong Ma,Xianchao Sun
Asparagine synthetase is a key enzyme that catalyses the conversion of amide groups from glutamine or ammonium to aspartate, which leads to the generation of asparagine. However, the role of asparagine synthetase in plant immunity remains largely unknown. Here, we identified a Nicotiana benthamiana asparagine synthetase B (NbAS-B) that associates with tomato mosaic virus coat protein-interacting protein L (IP-L) using the yeast two-hybrid assay and examined its role in tobacco mosaic virus (TMV) resistance. The association of IP-L with NbAS-B was further confirmed by in vivo co-immunoprecipitation, luciferase complementation imaging, and bimolecular fluorescence complementation assays. IP-L and NbAS-B interact in the nucleus and cytosol and IP-L apparently stabilizes NbAS-B, thus enhancing its accumulation. The expressions of IP-L and NbAS-B are continuously induced on TMV-green fluorescent protein (GFP) infection. Co-silencing of IP-L and NbAS-B facilitates TMV-GFP infection. Overexpression of NbAS-B in tobacco reduces TMV-GFP infection by significantly improving the synthesis of asparagine. Furthermore, the external application of asparagine significantly inhibits the infection of TMV-GFP by activating the salicylic acid signalling pathway. These findings hold the potential for the future application of asparagine in the control of TMV.
研究领域:
植物病理学
·
分子生物学
·
植物科学
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DOI
PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2024
·
IF 5.7
Zilin Wu,Min Huang,Junmei Jiang,Chun Zhang,Gefei Hao,Moxian Chen,Qing X. Li,Mengao Jia,Junfeng Liu,Xiangyang Li
Ningnanmycin has been proven to effectively inhibit infection by potato virus Y (PVY), although its underlying mechanisms remain unclear. In this study, we report a novel finding that ningnanmycin affects PVY replication. Two approaches were employed: studies using PVY movement-deficient mutants suggest that ningnanmycin, at a concentration of 500 μg/mL, inhibits PVY replication. Ningnanmycin interacts with the PVY-encoded coat protein (CP) with a binding constant of 1.34 μmol/L, and key amino acids Glu168 and Thr206 are involved in this interaction. Additionally, ningnanmycin induces the expression of antiviral response genes in Nicotiana benthamiana, including PRXIIB, PRXIIE, PUB4, and PER42. Furthermore, studies revealed that the overexpression of PUB4 in N. benthamiana confers resistance to PVY infection. These findings highlight the mechanisms by which ningnanmycin activates the PUB4 gene and suppresses CP assembly, thereby inhibiting PVY in N. benthamiana. This study represents an important step toward elucidating the molecular mechanism underlying the antiviral activity of ningnanmycin.
研究领域:
植物病理学
·
病毒学
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分子生物学
·
植物生物技术
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产品关联
DOI
PubMed
高影响因子
Current Plant Biology
·
2025
·
IF 4.5
Anastasia Barsukova,Lei Zhao,Svetlana Trusova
Phytaspases are specific plant proteases related to stress development and plant defense responses to adverse factors, such as insect or agrobacterial attacks. Phytaspases belong to the subtilisin-like proteases (subtilases), and they are distinct from other subtilases due to their strict substrate specificity and unusual life cycle. Phytaspases are localized in the apoplast under normal circumstances, sharing this feature with the majority of other subtilases. However, phytaspases move into the cell interior under stress development via a clathrin-mediated mechanism, and among subtilases, such behavior has been described only for phytaspases. The unique specificity in the structure of phytaspases enables in silico genomic analysis to predict candidates for phytaspases among the numerous plant subtilases. For example, three subtilases have been expected to be phytaspases for the plant Nicotiana benthamiana . Here, we demonstrate that N. benthamiana possesses a phytaspase-like activity that alters localization upon stress induction, and we isolate and purify the enzyme corresponding to this activity from leaf apoplast washes. Mass spectrometric analysis is used to identify the purified enzyme and establish which protein corresponds to this activity. Additionally, we characterize the substrate preferences of the N. benthamiana phytaspase.
研究领域:
植物学
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植物生物化学
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分子植物-微生物互作
·
蛋白质组学
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产品关联
DOI
高影响因子
BIOTECHNOLOGY LETTERS
·
2022
·Vol. 45
·期 1
·115-124
·
IF 2.716
Jin Jing-Hao,Zhou Xue,Liu Wang,Zhang Zi-Hui,Huang Shen-Xin,Zhao Wen-Jing,Chen Xiao-Ren
Objective To examine the influence of widely used protein affinity tags and the tobacco PR1a signal peptide (SP) on detection, purification and bioactivity analyses of the small oomycete apoplastic effector SCR96 in planta . Results Through agroinfiltration, the phytotoxic effector SCR96 of Phytophthora cactorum was expressed in Nicotiana benthamiana leaf apoplast as a fusion protein carrying single affinity tag (His, HA or FLAG) at either C- or N-terminus. Leaf necrosis caused by different affinity-tagged SCR96 varied among tags and replicates. All of tagged proteins can be detected by antibodies against SCR96. All of SCR96 fusions except N-terminally fused 6His-tagged protein were detected using tag antibodies, indicating that 6His tag may be degraded when fused at N-terminus. Interestingly, C-terminal His- and FLAG-tagged SCR96 maintained the biological activity after purification. In the substitution assay of SCR96 SP, we observed that PR1a SP can lead chimeric SCR96 expression in N. benthamiana , but the replacement totally disrupted its bioactivity. Conclusion C-terminal His or FLAG tag, along with its original SP, is efficient enough to enable detection and purification of functional SCR96 from N. benthamiana leaf apoplast, which would facilitate plant–pathogen interaction studies.
研究领域:
植物病理学
·
分子生物学
·
蛋白质工程
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产品关联
DOI
PubMed
高影响因子
RSC Advances
·
2017
·Vol. 7
·期 34
·21028-21035
·
IF 3.108
Yuyu Liu,Xiaohui Wang,Ting Mo,Yaru Yan,Yuelin Song,Yunfang Zhao,Jun Li,Shepo Shi,Xiao Liu,Pengfei Tu
Recently, owing to the important pharmaceutical properties of malonylated glycosides, their chemoenzymatic synthesis using malonyltransferase has received significant attention. In the present study, a new malonyltransferase, NbMaT1, was identified from Nicotiana benthamiana. Extensive enzymatic assays revealed its significant substrate tolerance based on HPLC-UV and HR-MS analyses. Moreover, 16 of the tested glycosides including flavone glycosides, flavonol glycosides, dihydroflavone glycosides, isoflavone glycosides, coumarin glycosides, and phenylethyl chromone glycosides with various sugar moieties (such as glucose, xylose, and galactose) substituted at different positions of their skeleton could be accepted by NbMaT1 to conduct the corresponding malonylation. Among these, enzymatic malonylation of phenylethyl chromone glycosides as well as xylosides and galactosides has rarely been reported earlier. Furthermore, one-pot synthesis using the known malonyl-CoA synthetase MatB and NbMaT1 as well as an unnatural fusion protein MatB–NbMaT1 was designed, which allowed malonic acid to be directly used in the malonylation reaction without the addition of expensive malonyl-CoA. Moreover, a remarkably improved conversion rate was observed for all the tested substrates, with both commercial and industrial application values. The malonylated product of the bioactive flavonoid diglycoside icariin was prepared and NMR spectroscopy revealed that the malonyl group was specifically transferred onto the 6-OH group of the glucose moiety. NbMaT1 was expected to be a universal and effective tool for chemoenzymatic synthesis of diverse bioactive-malonylated glycoside derivatives for drug discovery.
研究领域:
生物化学
·
药理学
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植物生物技术
·
酶学
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DOI
高影响因子
STAR Protocols
·
2021
·
IF 0
Yunjing Wang,Qian Gong,Fan Huang,Linfang He,Yule Liu
Summary Wounding evokes transient increases in cytosolic calcium (Ca 2+ ) concentration. Visualizing real-time Ca 2+ flux provides new insights into Ca 2+ -signaling pathways. Here, we outline a protocol to detect insect feeding-induced Ca 2+ flux elevation in Nicotiana benthamiana leaves based on the GCaMP3 reporter system by Leica fluorescence stereo microscopes (LFSM). LFSM combines super-fast manual screening with high-end imaging capabilities. Through this protocol, we can clearly observe the calcium flow after aphid’s piercing-sucking. Additionally, we describe a protocol to quantify Ca 2+ level using LFSM. For complete details on the use and execution of this protocol, please refer to Wang et al. (2021) .
研究领域:
植物生物学
·
分子生物学
·
昆虫学
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DOI
PubMed
PLANT CELL AND ENVIRONMENT
·
2024
·
IF 6
Jun-Xia Wang,Wen-Hao Han,Rui Xie,Feng-Bin Zhang,Zhi-Wei Ge,Shun-Xia Ji,Shu-Sheng Liu,Xiao-Wei Wang
Nicotiana benthamiana , a widely acknowledged laboratory model plant for molecular studies, exhibits lethality to certain insect pests and can serve as a dead-end trap plant for pest control in the field. However, the underlying mechanism of N. benthamiana 's resistance against insects remains unknown. Here, we elucidate that the lethal effect of N. benthamiana on the whitefly Bemisia tabaci arises from the toxic glandular trichome exudates. By comparing the metabolite profiles of trichome exudates, we found that 51 metabolites, including five O -acyl sugars ( O -AS) with medium-chain acyl moieties, were highly accumulated in N. benthamiana . Silencing of two O -AS biosynthesis genes, branched-chain keto acid dehydrogenase ( BCKD ) and Isopropyl malate synthase-C ( IPMS-C ), significantly reduced the O -AS levels in N. benthamiana and its resistance against whiteflies. Additionally, we demonstrated that the higher expression levels of BCKD and IPMS-C in the trichomes of N. benthamiana contribute to O -AS synthesis and consequently enhance whitefly resistance. Furthermore, overexpression of NbBCKD and NbIPMS-C genes in the cultivated tobacco Nicotiana tabacum enhanced its resistance to whiteflies. Our study revealed the metabolic and molecular mechanisms underlying the lethal effect of N. benthamiana on whiteflies and presents a promising avenue for improving whitefly resistance.
研究领域:
植物科学
·
分子生物学
·
昆虫学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Phytopathology Research
·
2022
·Vol. 4
·期 1
·1-12
·
IF 3.955
Jiang Long,Xiang Shunyu,Lv Xing,Wang Xiaoyan,Li Fengwei,Liu Weina,Liu Changyun,Ran Mao,Huang Jin,Xu Xiaohong,Ma Xiaozhou,Jin Yabo,Sun Xianchao
Silver (Ag)-containing agents or materials are widely used today in plant protection for their antimicrobial activity. In view of the superior inhibitory ability of biosynthesized (aldehyde-modified sodium alginate based) silver nanoparticles (AgNPs) against plant pathogenic fungi in our previous research, here we explored the antagonistic effect of biosynthesized AgNPs on plant pathogenic bacteria and the underlying mechanism. We selected Pseudomonas syringae pv. tabaci , the causal agent of tobacco wildfire disease, as the target and found that 1.2 μg/mL biosynthesized AgNPs completely inhibited the growth of P. syringae pv. tabaci in vitro and in vivo by partly destroying the cell membrane structure of the pathogen, resulting in cytoplasmic leakage. Moreover, Nicotiana benthamiana treated with 1.2 μg/mL biosynthesized AgNPs exhibited a significant upregulation of nonexpressor of pathogenesis-related genes 1 ( NPR1 ) and pathogenesis-related gene 2 ( PR2 ), the typical markers of the salicylic acid (SA)-mediated defense system, and an increase in peroxidase (POD) and polyphenol oxidase (PPO) activities as well as the production of reactive oxygen species (ROS). Furthermore, biosynthesized AgNPs treatment increased the chlorophyll content and dry weight of N. benthamiana . Overall, we demonstrated that biosynthesized AgNPs at a low concentration have high inhibitory effect on the pathogen causing tobacco wildfire disease by destroying bacterial cell membrane and inducing defense resistance in host plant. These results lay a theoretical foundation for further application of biosynthesized AgNPs in the control of plant bacterial diseases.
研究领域:
植物病理学
·
纳米技术
·
分子植物-微生物互作
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DOI
高影响因子
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
·
2025
·Vol. 26
·期 3
·1384
·
IF 4.9
Sadia Noorin,Youwei Du,Yi Liu,Shuanghong Wang,Yan Wang,Hongchen Jia,Tom Hsiang,Rong Zhang,Guangyu Sun
Potassium (K) fertilization is crucial for plant resistance to pathogens, but the underlying mechanisms remain unclear. Here, we investigate the molecular mechanism by which the addition of K promotes resistance inNicotiana benthamianatoPhytophthora parasitica. We found thatN. benthamianawith high K content (HK, 52.3 g/kg) produced more ethylene in response toP. parasiticainfection, compared toN. benthamianawith low-K content (LK, 22.4 g/kg). An exogenous ethylene application effectively increased resistance in LKN. benthamianato the level under HK status, demonstrating the involvement of ethylene in the HK-associated resistance inN. benthamiana. Further, transcriptome analysis showed thatNbSAMS1, encoding ethylene biosynthesis, was induced to upregulateP. parasiticaabout five times higher in HK than in LKN. benthamiana. NbSAMS1overexpression enhanced resistance in LK plants, whereasNbSAMS1silencing reduced resistance in HK plants, confirming its importance in conferring resistance. Furthermore, we identified a calcium-binding protein, NbCBP1, which interacted with NbSAMS1, promoting its expression in HKN. benthamiana.SilencingNbCBP1compromised resistance in HKN. benthamiana, whereas its overexpression improved resistance in LKN. benthamiana. Notably, NbCBP1 protected NbSAMS1 from degradation by the 26S proteasome, thereby sustaining ethylene accumulation in HKN. benthamianain response toP. parasiticainfection. Thus, our research elucidated some mechanisms of the NbCBP1-NbSAMS1 module associated with disease resistance in HKN. benthamiana.
研究领域:
植物病理学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
PLANTA
·
2022
·Vol. 256
·期 5
·1-11
·
IF 4.54
Yang Daoyong,Peng Qiding,Cheng Yongchao,Xi Dehui
Main conclusion G6PDH negatively regulates viral accumulation in Nicotiana benthamiana through RBOHB-associated ROS signaling. Anti-oxidative metabolism and phytohormone-mediated immunity responses play important roles in virus infection. Abstract Glucose-6-phosphate dehydrogenase (G6PDH) is an enzyme in the pentose phosphate pathway, which plays an important role in maintaining intracellular redox homeostasis and has functions in plant growth, development and stress tolerance. However, the role of G6PDH in plants response to virus infection is poorly understood. In this study, NbG6PDH was found to be down-regulated after Chilli veinal mottle virus (ChiVMV-GFP) infection in Nicotiana benthamiana . Subcellular localization of NbG6PDH showed that it was punctate distributed in the protoplasm. Silencing of NbG6PDH reduced the sensitivity of N. benthamiana plants to ChiVMV-GFP. By contrast, transient overexpression of NbG6PDH promoted the accumulation of the virus. The results of physiological indexes showed that glutathione (GSH), catalase (CAT) and proline played an important role in maintaining plants physiological homeostasis. The results of gene expression detection showed that jasmonic acid/ethylene (JA/ET) signaling pathway was significantly correlated with the response of N. benthamiana to ChiVMV-GFP infection, and the changes of N. benthamiana respiratory burst oxidase homologues B (NbRBOHB) indicated that the NbG6PDH-dependent ROS may be regulated by NbRBOHB. Pretreatment of the inducer of reactive oxygen species (ROS) promoted virus infection, whereas inhibitor of ROS alleviated virus infection. Thus, our results indicate that the promoting effect of NbG6PDH on ChiVMV-GFP infection may be related to the NbRBOHB-regulated ROS production.
研究领域:
植物病毒学
·
分子植物生理学
·
植物病理学
✓ 已验证
DOI
PubMed
高影响因子
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