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1.
Expression of the ipomoelin gene from sweet potato is regulated by dephosphorylated proteins, calcium ion and ethylene 总被引:2,自引:1,他引:1
Y.-C. CHEN B.-W. TSENG Y.-L. HUANG Y.-C. LIU & S.-T. JENG 《Plant, cell & environment》2003,26(8):1373-1383
A wound‐inducible cDNA, ipomoelin (IPO) was isolated from the subtraction library of sweet potato (Ipomoea batatas cv. Tainung 57) and used as a molecular probe to investigate the transduction pathway of wounding signal within plant cells. Following mechanical wounding of the leaves of sweet potato, IPO mRNA accumulation peaked at 6 h and then continuously declined. However, IPO gene expression in the apical unwounded leaves began at 6 h after wounding and continued for a further 10 h. Besides mechanical wounding, methyl jasmonate (MeJA) was identified as a signal transducer leading to the accumulation of IPO mRNA. Treatment with salicylic acid reduced the production of IPO mRNA, further supporting the involvement of the octadecanoid pathway in the signal transduction of wounding in sweet potato. In addition, ethylene was involved in the signal pathway and induced the expression of the IPO gene. Furthermore, the application of okadaic acid, a protein phosphatase inhibitor, blocked the accumulation of IPO mRNA induced by MeJA or ethylene, indicating that activation of the IPO gene by both MeJA and ethylene was via dephosphorylated proteins. The presence of a calcium ion chelator or channel blockers also inhibited the expression of the IPO gene after wounding. However, investigation by confocal scanning microscopy further pointed out that mechanical wounding rather than the application of MeJA induced the accumulation of the calcium ion. These results may indicate that the calcium ion is also involved in the activation of IPO mRNA. In addition, wounding signals the accumulation of calcium ion first and then stimulates the biosynthesis of MeJA in sweet potato. Hence, the reaction sequence of signal transducers, including the calcium ion, MeJA and protein kinase/phosphatase, in the wounding signalling pathway of sweet potato is suggested in this report. 相似文献
2.
Involvement of hydrogen peroxide and nitric oxide in expression of the ipomoelin gene from sweet potato 总被引:9,自引:0,他引:9
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The IPO (ipomoelin) gene was isolated from sweet potato (Ipomoea batatas cv Tainung 57) and used as a molecular probe to investigate its regulation by hydrogen peroxide (H(2)O(2)) and nitric oxide (NO) after sweet potato was wounded. The expression of the IPO gene was stimulated by H(2)O(2) whether or not the plant was wounded, but its expression after wounding was totally suppressed by the presence of diphenylene iodonium, an inhibitor of NADPH oxidase, both in the local and systemic leaves of sweet potato. These results imply that a signal transduction resulting from the mechanical wounding of sweet potato may involve NADPH oxidase, which produces endogenous H(2)O(2) to stimulate the expression of the IPO gene. The production of H(2)O(2) was also required for methyl jasmonate to stimulate the IPO gene expression. On the contrary, NO delayed the expression of the IPO gene, whereas N(G)-monomethyl-L-arginine monoacetate, an inhibitor of NO synthase, enhanced the expression of the IPO gene after the plant was wounded. This study also demonstrates that the production of H(2)O(2) stained with 3,3'-diaminobenzidine hydrochloride could be stimulated by wounding but was suppressed in the presence of NO. Meanwhile, the generation of NO was visualized by confocal scanning microscope in the presence of 4,5-diaminofluorescein diacetate after sweet potato was wounded. In conclusion, when sweet potato was wounded, both H(2)O(2) and NO were produced to modulate the plant's defense system. Together, H(2)O(2) and NO regulate the expression of the IPO gene, and their interaction might further stimulate plants to protect themselves from invasions by pathogens and herbivores. 相似文献
3.
Shiyang Li Xue-Ao Liu Lizi Zhao Huiqing Huang Bei Li Zhizhong Song Meixia Liang Hongxia Zhang Limin Wang Shenglin Zhou 《Phyton》2021,90(3):761-772
Seven in absentia (SINA) family proteins play a central role in plant growth, development and resistance to abiotic stress. However, their biological function in plant response to cold stress is still largely unknown. In this work, a seven in absentia gene IbSINA5 was isolated from sweet potato. Quantitative real-time polymerase chain reaction (qRT-PCR) analyses demonstrated that IbSINA5 was ubiquitously expressed in various tissues and organs of sweet potato, with a predominant expression in fibrous roots, and was remarkably induced by cold, drought and salt stresses. Subcellular localization assays revealed that IbSINA5-GFP fusion protein was mainly localized in cytoplasm and nucleus. Overexpression of IbSINA5 in sweet potato led to dramatically improved resistance to cold stress in transgenic plants, which was associated with the up-regulated expression of IbCOR (cold-regulated) genes, increased proline production, and decreased malondialdehyde (MDA) and H2O2 accumulation in the leaves of transgenic plants. Furthermore, transient expression of IbCBF3, a C-repeat binding factor (CBF) gene, in the leaf protoplasts of wild type sweet potato plants up-regulated the expression of both IbSINA5 and IbCOR genes. Our results suggest that IbSINA5 could function as a positive regulator in the cold signaling pathway through a CBF-SINA-COR mediated module in sweet potato, and have a great potential to be used as a candidate gene for the future breeding of new plant species with improved cold resistance. 相似文献
4.
Zhitong Ren Shaozhen He Ning Zhao Hong Zhai Qingchang Liu 《Plant biotechnology journal》2019,17(1):21-32
Sucrose non‐fermenting‐1‐related protein kinase‐1 (SnRK1) is an essential energy‐sensing regulator and plays a key role in the global control of carbohydrate metabolism. The SnRK1 gene has been found to increase starch accumulation in several plant species. However, its roles in improving starch quality have not been reported to date. In this study, we found that the IbSnRK1 gene was highly expressed in the storage roots of sweet potato and strongly induced by exogenous sucrose. Its expression followed the circandian rhythm. Its overexpression not only increased starch content, but also decreased proportion of amylose, enlarged granule size and improved degree of crystallinity and gelatinization in transgenic sweet potato, which revealed, for the first time, the important roles of SnRK1 in improving starch quality of plants. The genes involved in starch biosynthesis pathway were systematically up‐regulated, and the content of ADP‐glucose as an important precursor for starch biosynthesis and the activities of key enzymes were significantly increased in transgenic sweet potato. These findings indicate that IbSnRK1 improves starch content and quality through systematical up‐regulation of the genes and the increase in key enzyme activities involved in starch biosynthesis pathway in transgenic sweet potato. This gene has the potential to improve starch content and quality in sweet potato and other plants. 相似文献
5.
Chengyang Li Lianjun Wang Shasha Chai Yinghua Xu Chong Wang Yi Liu Jian Lei Xiaojie Jin Xianliang Cheng Yuanyuan Yang Xinsun Yang Wenying Zhang 《Phyton》2022,91(8):1779-1793
Fusarium wilt, a disease caused by Fusarium oxysporum f.sp batatas (Fob) is an important disease in sweet potato production. Using endophytic bacteria for biological control of sweet potato diseases is one of the important ways. A Bacillus subtilis with antagonistic effect on Fusarium wilt of sweet potato was isolated from soil by confrontation culture. According to the biological characteristics, 16S rDNA sequence analysis, and physiological and biochemical analysis, the Bacillus subtilis HAAS01 was named. A pot experiment was conducted for the biological control experiment of strain HAAS01, and the endogenous hormone content, antioxidant enzyme activity, soluble protein content, and related gene expressions of sweet potato plants were detected. The results showed that the HAAS01 strain could promote the production of endogenous hormones and resist the infection of plant diseases together with defensive enzymes and upregulation of related gene expressions. In summary, Bacillus subtilis HAAS01 was effective in controlling Fusarium wilt of sweet potato and has potential for application and development. 相似文献
6.
Inhibition of the gene expression for granule-bound starch synthase I by RNA interference in sweet potato plants 总被引:3,自引:0,他引:3
Otani M Hamada T Katayama K Kitahara K Kim SH Takahata Y Suganuma T Shimada T 《Plant cell reports》2007,26(10):1801-1807
Granule-bound starch synthase I (GBSSI) is one of the key enzymes catalyzing the formation of amylose, a linear α(1,4)D-glucan
polymer, from ADP-glucose. Amylose-free transgenic sweet potato plants were produced by inhibiting sweet potato GBSSI gene expression through RNA interference. The gene construct consisting of an inverted repeat of the first exon separated
by intron 1 of GBSSI driven by the CaMV 35S promoter was integrated into the sweet potato genome by Agrobacterium tumefaciens-mediated transformation. In over 70% of the regenerated transgenic plants, the expression of GBSSI was inactivated giving rise to storage roots containing amylopectin but not amylose. Electrophoresis analysis failed to detect
the GBSSI protein, suggesting that gene silencing of the GBSSI gene had occurred. These results clearly demonstrate that amylose synthesis is completely inhibited in storage roots of sweet
potato plants by the constitutive production of the double-stranded RNA of GBSSI fragments. We conclude that RNA interference is an effective method for inhibiting gene expression in the starch metabolic
pathway. 相似文献
7.
Sporamin, a tuberous storage protein of sweet potato, was systemically expressed in leaves and stems by wound stimulation. In an effort to demonstrate the regulatory mechanism of wound response on the sporamin gene, a 1.25 kb sporamin promoter was isolated for studying the wound-induced signal transduction. Two wound response-like elements, a G box-like element and a GCC core-like sequence were found in this promoter. A construct containing the sporamin promoter fused to a -glucuronidase (GUS) gene was transferred into tobacco plants by Agrobacterium-mediated transformation. The wound-induced high level of GUS activity was observed in stems and leaves of transgenic tobacco, but not in roots. This expression pattern was similar to that of the sporamin gene in sweet potatoes. Exogenous application of methyl jasmonate (MeJA) activated the sporamin promoter in leaves and stems of sweet potato and transgenic tobacco plants. A competitive inhibitor of ethylene (2,5-norbornadiene; NBD) down-regulated the effect of MeJA on sporamin gene expression. In contrast, salicylic acid (SA), an inhibitor of the octadecanoid pathway, strongly suppressed the sporamin promoter function that was stimulated by wound and MeJA treatments. In conclusion, wound-response expression of the sporamin gene in aerial parts of plants is regulated by the octadecanoid signal pathway. 相似文献
8.
Roderick Lawton Stacy Winfield Hanery Daniell Ajmer S. Bhagsari Sarwan K. Dhir 《Plant Molecular Biology Reporter》2000,18(2):139-139
Green-fluorescent protein (GFP) gene expression, transient and stable after electroporation and particle bombardment, was
analyzed in tissues of sweet potato cv.Beauregard. Leaf and petiole tissues were used for protoplast isolation and electroporation. After 48 h, approximately 25–30% of electroporated
mesophyll cell protoplasts regenerated cell walls, and of these, 3% expressed GFP. Stable expression of GFP after four weeks
of culture was observed in 1.0% of the initial GFP positive cells. In a separate experiment, we observed 600–700 loci expressing
GFP 48 h after bombarding leaf tissue or embryogenic calli, and stable GFP-expressing sectors were seen in leaf-derived embryogenic
calli after four weeks of protoplast culture without selection. These results demonstrate GFP gene expression in sweet potato
tissues. Screening for GFP gene expression may prove useful to improve transformation efficiency and to facilitate detection
of transformed sweet potato plants. 相似文献
9.
10.
A gene encoding the outer capsid glycoprotein (VP7) of simian rotavirus SA11, was genetically linked to the amino terminus
of the ricin toxin B subunit (RTB) isolated from castor-oil plant (Ricinus communis) seeds. To assess fusion protein expression in plant cells, the VP7::RTB fussion gene was transferred into potato (Solanum tuberosum) cells by Agrobacterium tumefaciens-mediated transformation methods and transformed plants regenerated. The fusion gene was detected in transformed potato genomic
DNA by polymerase chain reaction DNA amplification methods. Immunoblot analysis with anti-SA11 antiserum as the primary antibody
verified the presence of VP7::RTB fusion protein in transformed potato tuber tissues. The plant-synthesized fusion protein
bound RTB membrane receptors as measured by asialofetuin-enzyme-linked immunosorbent assay (ELISA). The ELISA results indicated
that the VP7::RTB fusion protein was biologically active and made up approx 0.03% of total soluble transformed tuber protein.
The biosynthesis of receptor binding VP7::RTB fusion protein in potato tissues demonstrates the feasibility of producing monomeric
ricin toxin B subunit adjuvant-virus antigen fusion proteins in crop plants for enhanced immunity. 相似文献
11.
12.
Overexpression of the IbMYB1 gene in an orange‐fleshed sweet potato cultivar produces a dual‐pigmented transgenic sweet potato with improved antioxidant activity
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Sung‐Chul Park Yun‐Hee Kim Sun Ha Kim Yu Jeong Jeong Cha Young Kim Joon Seol Lee Ji‐Yeong Bae Mi‐Jeong Ahn Jae Cheol Jeong Haeng‐Soon Lee Sang‐Soo Kwak 《Physiologia plantarum》2015,153(4):525-537
13.
Herbicide-resistant sweet potato plants were produced through biolistics of embryogenic calli derived from shoot apical meristems.
Plant materials were bombarded with the vectors containing the β-glucuronidase gene (gusA) and the herbicide-resistant gene (bar). Selection was carried out using phosphinothricin (PPT). Transformants were screened by the histochemical GUS and Chlorophenol
Red assays. PCR and Southern-blot analyses indicated the presence of introduced bar gene in the genomic DNA of the transgenic plants. When sprayed with Basta, the transgenic sweet potato plants was tolerant
to the herbicide. Hence, we report successful transformation of the bar gene conferring herbicide resistance to sweet potato. 相似文献
14.
15.
16.
Katsutada Takahashi 《Bioscience, biotechnology, and biochemistry》2013,77(7):629-633
In the latest work, a method to determine calorimetrically the α-glucosidic linkage contents in starches was introduced.The present work refers to the application of this method to the determination of the α-1,4 glucosidic linkage content in two kinds of sweet potato starch (i.e., Norin-1 and Okinawa-100) isolated at the two different stages of development.It has been found that the sample isolated on October 31 (1953) had a larger value for α-1,4 glucosidic linkage content than the sample isolated on August 6 (1953) for both the two sweet potato starches.Combined with the amperometric titration method the increase in α-1,4 glucosidic linkage content has been found to be due to both the increases in amylose content and in average unit chain length of amylopectin component. 相似文献
17.
以超表达甘薯橙色基因(IbOr)的转基因甘薯(TS)以及非转基因甘薯(NT)为实验材料,通过15%聚乙二醇6000(PEG-6000)模拟干旱条件,研究转基因与非转基因甘薯幼苗在水分胁迫不同时间的光合系统,膜脂过氧化及抗氧化防御系统中主要指标的变化情况,探讨转基因甘薯耐旱性的生理机制。结果显示:(1)随PEG-6000胁迫时间延长,甘薯叶片的叶绿素、类胡萝卜素含量及其叶片净光合速率、气孔导度、胞间CO2浓度、蒸腾速率都显著降低,但转基因株系降低幅度小于非转基因植株。(2)在正常供水和水分胁迫下,超表达IbOr基因甘薯叶片中O-·2、MDA含量均低于非转基因甘薯,即转基因甘薯具有较低的活性氧水平且脂膜受损伤较小。(3)PEG-6000胁迫24h后,甘薯叶片中SOD、POD酶活性均增加,48h达到最大值,且转基因甘薯中2种酶活性显著高于非转基因甘薯。研究表明,过表达IbOr基因可以有效减轻甘薯在水分胁迫条件下受损害的程度,且可能主要通过提高甘薯的抗氧化胁迫能力来完成。 相似文献
18.
Sweet potato NAC transcription factor,IbNAC1, upregulates sporamin gene expression by binding the SWRE motif against mechanical wounding and herbivore attack
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Shi‐Peng Chen I Winnie Lin Xuanyang Chen Yin‐Hao Huang Shiao‐Chi Chang Hui‐Shan Lo Hseuh‐Han Lu Kai‐Wun Yeh 《The Plant journal : for cell and molecular biology》2016,86(3):234-248
19.
A gene encoding VP7, the outer capsid protein of simian rotavirus SA11, was fused to the carboxyl terminus of the cholera
toxin B subunit gene. A plant expression vector containing the fusion gene under control of the mannopine synthase P2 promoter was introduced into Solanum tuberosum cells by Agrobacterium tumefaciens-mediated transformation. The CTB::VP7 fusion gene was detected in the genomic DNA of transformed potato leaf cells by polymerase chain reaction (PCR) amplification
methods. Immunoblot analysis of transformed potato tuber tissue extracts showed that synthesis and assembly of the CTB::VP7
fusion protein into oligomers of pentameric size occurred in the transformed plant cells. The binding of CTB::VP7 fusion protein
pentamers to sialo-sugar containing GM1 ganglioside receptors on the intestinal epithelial cell membrane was quantified by
enzyme-linked immunosorbent assay (ELISA). The ELISA results showed that the CTB::VP7 fusion protein made up approx 0.01%
of the total soluble tuber protein. Synthesis and assembly of CTB::VP7 monomers into biologically active pentamers in transformed
potato tubers demonstrates the feasibility of using edible plants as a mucosal vaccine for the production and delivery system
for rotavirus capsid protein antigens. 相似文献
20.
Stacy Winfield Rodrick Lawton Henry Daniell Sarwan K. Dhir 《In vitro cellular & developmental biology. Plant》2001,37(5):648-653
Summary The expression of the green-fluorescent protein (GFP) gene from Aequorea victoria (jellyfish) was analyzed by transient and stable expression in sweet potato Ipomoea batatas L. (Lam.) ev. Beauregard tissues by electroporation and particle bombardment. Leaf and petiole segments from in vitro-raised young plantlets were used for protoplast isolation and electroporation. Embyrogenic callus was also produced from
leaf segments for particle bombardment experiments. A buffer solution containing 1×106 protoplasts ml−1 was mixed with plasmid DNA containing the GFP gene, and electroporated at 375 V cm−1. Approximately 25–30% of electroporated mesophyll cell protoplasts subsequently cultured in KM8P medium regenerated cell
walls after 48 h. Of these, 3% emitted bright green fluorescence when exposed to UV-blue light at 395 nm. Transformed cells
continued to grow after embedding in KM8P medium solidifed with 1.2% SeaPlaque agarose. Stable expression of GFP was observed
after 4 wk of culture in approximately 1.0% of the initial GFP positive cells (27.5 GFP positive micro callases out of 3024
cells which transiently expressed GFP 48 h after electroporation). In a separate experiment, 600–700 bright green spots were
observed per plate 48 h after bombarding leaf segments or embryogenic cellus. In bombarded cultures, several stable GEP-expressing
sectors were observed in leafderived embryogenic callus grown without selection for 4 wk. These results show that GFP gene
expression can occur in various sweet potato tissues, and that it may be a useful sereenable marker to improve transformation
efficiency and obtain transgenic sweet potato plants. 相似文献