WY195, a New Inducible Promoter From the Rubber Powdery Mildew Pathogen, Can Be Used as an Excellent Tool for Genetic Engineering

Until now, there are few studies and reports on the use of endogenous promoters of obligate biotrophic fungi. The WY195 promoter in the genome of Oidium heveae, the rubber powdery mildew pathogen, was predicted using PromoterScan and its promoter function was verified by the transient expression of the β-glucuronidase (GUS) gene. WY195 drove high levels of GUS expression in dicotyledons and monocotyledons. qRT-PCR indicated that GUS expression regulated by the WY195 promoter was 17.54-fold greater than that obtained using the CaMV 35S promoter in dicotyledons (Nicotiana tabacum), and 5.09-fold greater than that obtained using the ACT1 promoter in monocotyledons (Oryza sativa). Furthermore, WY195-regulated GUS gene expression was induced under high-temperature and drought conditions. Soluble proteins extracted from WY195-hpaXm transgenic tobacco was bioactive. Defensive micro-HR induced by the transgene expression of hpaXm was observed on transgenic tobacco leaves. Disease resistance bioassays showed that WY195-hpaXm transgenic tobacco enhanced the resistance to tobacco mosaic virus (TMV). WY195 has great potential for development as a new tool for genetic engineering. Further in-depth studies will help to better understand the transcriptional regulation mechanisms and the pathogenic mechanisms of O. heveae.

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WY195, a new inducible promoter from the rubber powdery mildew pathogen, can be used as an excellent tool for genetic engineering

10.21203/rs.2.19804/v1 ◽

2020 ◽

Author(s):

Yi Wang ◽

Mamy Jayne Nelly Rajaofera ◽

Li Zhu ◽

Jinyao Yin ◽

Chen Wang ◽

...

Keyword(s):

Powdery Mildew ◽

Genetic Engineering ◽

Transgenic Tobacco ◽

Inducible Promoter ◽

35S Promoter ◽

Inducible Promoters ◽

Organ Tissue ◽

Hylocereus Undatus ◽

Powdery Mildew Pathogen ◽

Excellent Tool

Abstract Background: In recent years, the rational use of inducible promoters and organ/tissue-specific promoters have become a focus of promoter research because these enable greater control of expression.Results: The WY195 promoter in the genome of Oidium heveae, the rubber powdery mildew pathogen, was predicted using PromoterScan and its promoter function was verified by the transient expression of the b-glucuronidase (GUS) gene. WY195 drove high levels of GUS expression in dicotyledons (Nicotiana tabacum, Hevea brasiliensis Reyan 7-33-97 and Hylocereus undatus) and monocotyledons (Oryza sativa ssp. japonica cv. Nipponbare, Hordeum vulgare and Zea mays). qRT-PCR indicated that GUS expression regulated by the WY195 promoter was 17.54-fold greater than that obtained using the CaMV 35S promoter in dicotyledons (N. tabacum), and 5.09-fold greater than that obtained using the ACT1 promoter in monocotyledons (O. sativa). Furthermore, WY195-regulated GUS gene expression was induced under high-temperature and drought conditions. Soluble proteins extracted from WY195-hpaXm transgenic tobacco was bioactive. Defensive micro-HR induced by the transgene expression of hpaXm was observed on transgenic tobacco leaves. Disease resistance bioassays showed that WY195-hpaXm transgenic tobacco enhanced the resistance to tobacco mosaic virus (TMV).Conclusions: WY195 is an excellent endogenous inducible promoter with great potential for development as a new tool for genetic engineering and should also promote O. heveae molecular research, which to date has been a challenge owing to the obligate parasitic nature of this pathogen.

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Co-expression of AGPS and AGPL genes encoded for AGPase from cassava to enhance starch accumulation in transgenic tobacco

Vietnam Journal of Biotechnology ◽

10.15625/1811-4989/14/2/9354 ◽

2016 ◽

Vol 14 (2) ◽

pp. 287-293

Author(s):

Nguyễn Văn Đoài ◽

Nguyễn Minh Hồng ◽

Lê Thu Ngọc ◽

Nguyễn Thị Thơm ◽

Nguyễn Đình Trọng ◽

...

Keyword(s):

Transgenic Tobacco ◽

Higher Plants ◽

Starch Content ◽

Genetically Modified Crops ◽

Starch Accumulation ◽

35S Promoter ◽

Effective Strategies ◽

Plant Expression Vector ◽

Transgenic Lines ◽

Cassava Varieties

The AGPase (ADP-Glucose pyrophosphorylase) is one of the ubiquitous enzymes catalyzing the first step in starch biosynthesis. It plays an important role in regulation and adjusts the speed of the entire cycle of glycogen biosynthesis in bacteria and starch in plants. In higher plants, it is a heterotetramer and tetrameric enzyme consisting two large subunits (AGPL) and two small subunits (AGPS) and encoded by two genes. In this paper, both AGPS and AGPL genes were sucessfully isolated from cassava varieties KM140 and deposited in Genbank with accession numbers KU243124 (AGPS) and KU243122 (AGPL), these two genes were fused with P2a and inserted into plant expression vector pBI121 under the control of 35S promoter. The efficient of this construct was tested in transgenic N. tabacum. The presence and expression of AGPS and AGPL in transgenic plants were confirmed by PCR and Western hybridization. The starch content was quantified by the Anthrone method. Transgenic plant analysis indicated that that two targeted genes were expressed simultaneously in several transgenic tobacco lines under the control of CaMV 35S promoter. The starch contents in 4 analyzed tobacco transgenic lines displays the increase 13-116% compared to WT plants. These results indicated that the co-expression of AGPS and AGPL is one of effective strategies for enhanced starch production in plant. These results can provide a foundation for developing other genetically modified crops to increase starch accumulation capacity.

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Cloning and Characterization of a Novel Wheat Glycoside Hydrolase Gene TaGlc2 Induced by Powdery Mildew Pathogen (Erysiphe graminis) Infection

ACTA AGRONOMICA SINICA ◽

10.3724/sp.j.1006.2009.00786 ◽

2009 ◽

Vol 35 (5) ◽

pp. 786-794

Author(s):

N PUDAKE Ramesh ◽

Ming-Ming XIN ◽

Yu-Jing YIN ◽

Chao-Jie XIE ◽

Zhong-Fu NI ◽

...

Keyword(s):

Powdery Mildew ◽

Glycoside Hydrolase ◽

Erysiphe Graminis ◽

Powdery Mildew Pathogen

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Transformation of the cucurbit powdery mildew pathogen Podosphaera xanthii by Agrobacterium tumefaciens

New Phytologist ◽

10.1111/nph.14297 ◽

2016 ◽

Vol 213 (4) ◽

pp. 1961-1973 ◽

Cited By ~ 18

Author(s):

Jesús Martínez‐Cruz ◽

Diego Romero ◽

Antonio Vicente ◽

Alejandro Pérez‐García

Keyword(s):

Agrobacterium Tumefaciens ◽

Powdery Mildew ◽

Podosphaera Xanthii ◽

Powdery Mildew Pathogen ◽

Cucurbit Powdery Mildew

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Genetic engineering for in vivo optical interrogation of neuronal responses to cell type-specific silencing

10.1101/2021.01.13.426508 ◽

2021 ◽

Author(s):

Firat Terzi ◽

Johannes Knabbe ◽

Sidney B. Cambridge

Keyword(s):

High Resolution ◽

Genetic Engineering ◽

Transgene Expression ◽

Neuronal Morphology ◽

Dependent Manner ◽

Cell Type ◽

Fluorescent Marker ◽

Genetic Perturbations ◽

Cell Type Specific

SummaryGenetic engineering of quintuple transgenic brain tissue was used to establish a low background, Cre-dependent version of the inducible Tet-On system for fast, cell type-specific transgene expression in vivo. Co-expression of a constitutive, Cre-dependent fluorescent marker selectively allowed single cell analyses before and after inducible, tet-dependent transgene expression. Here, we used this method for acute, high-resolution manipulation of neuronal activity in the living brain. Single induction of the potassium channel Kir2.1 produced cell type-specific silencing within hours that lasted for at least three days. Longitudinal in vivo imaging of spontaneous calcium transients and neuronal morphology demonstrated that prolonged silencing did not alter spine densities or synaptic input strength. Furthermore, selective induction of Kir2.1 in parvalbumin interneurons increased the activity of surrounding neurons in a distance-dependent manner. This high-resolution, inducible interference and interval imaging of individual cells (high I5, ‘HighFive’) method thus allows visualizing temporally precise, genetic perturbations of defined cells.

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