Transient virus-induced gene silencing of MaBAM9b efficiently suppressed starch degradation during postharvest banana fruit ripening

Abstract Background: Banana is a typical starch conversion fruit, and fruit ripening involves a process of fruit quality formation. To improve the nutritional value of banana, it is necessary to understand the genetic basis of the metabolic pathways that operate during fruit ripening processes. MaBAM9b is a key enzyme gene that might play an important role in starch degradation during the banana fruit ripening process. The identification of gene function by stable genetic transformation is time- and energy-consuming. Thus, developing an efficient and rapid method for functional identification is imperative. Virus-Induced Gene Silencing (VIGS) is a reverse-genetics method based on RNA-mediated antiviral plant defense that has been used to rapidly identify gene function in plants. Results: In this report, 0.5% iodine-potassium-iodide (I2-KI) staining for 150 s determined that a 1:3 ratio of TRV1: TRV2-MaBAM9b cultivated at an optical density of 600 nm (OD600) 0.8 at 30 mmHg for 30 sec and kept on Murashige & Skoog (MS) media for 5 d produced the best silencing results. Under these conditions, the total starch content was greatly increased, while the β-amylase activity, soluble sugar contents, and the expression of endogenous MaBAM9b were greatly decreased. Conclusions: The developed system is particularly useful for studying genes and networks for starch conversion in fruit, which alone would not produce a visual phenotype. This system will provide a platform for banana functional genomics and for banana fruit quality improvement.

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Silencing of the phytoene desaturase (PDS) gene affects the expression of fruit-ripening genes in tomatoes

Plant Methods ◽

10.1186/s13007-019-0491-z ◽

2019 ◽

Vol 15 (1) ◽

Cited By ~ 1

Author(s):

Aung Htay Naing ◽

Swum Yi Kyu ◽

Phyo Phyo Win Pe ◽

Kyeung Il Park ◽

Je Min Lee ◽

...

Keyword(s):

Gene Silencing ◽

Fruit Ripening ◽

Tomato Fruit ◽

Carotenoid Biosynthesis ◽

Past Research ◽

Ethylene Biosynthesis ◽

Phytoene Desaturase ◽

Virus Induced Gene Silencing ◽

Pale Yellow ◽

Ripening Genes

Abstract Background Past research has shown that virus-induced phytoene desaturase (PDS) gene silencing via agroinjection in the attached and detached fruit of tomato plants results in a pale-yellow fruit phenotype. Although the PDS gene is often used as a marker for gene silencing in tomatoes, little is known about the role of PDS in fruit ripening. In this study, we investigated whether the pepper PDS gene silenced endogenous PDS genes in the fruit of two tomato cultivars, Dotaerang Plus and Legend Summer. Results We found that the pepper PDS gene successfully silenced endogenous PDS in tomato fruit at a silencing frequency of 100% for both cultivars. A pale-yellow silenced area was observed over virtually the entire surface of individual fruit due to the transcriptional reduction in phytoene desaturase (PDS), zeta-carotene (ZDS), prolycopene isomerase (CrtlSO), and beta-carotene hydroxylase (CrtR-b2), which are the carotenoid biosynthesis genes responsible for the red coloration in tomatoes. PDS silencing also affected the expression levels of the fruit-ripening genes Tomato AGAMOUS-LIKE1 (TAGL1), RIPENING INHIBITOR (RIN), pectin esterase gene (PE), lipoxygenase (LOX), FRUITFULL1/FRUITFUL2 (FUL1/FUL2), and the ethylene biosynthesis and response genes 1-aminocyclopropane-1-carboxylate oxidase 1 and 3 (ACO1 and ACO3) and ethylene-responsive genes (E4 and E8). Conclusion These results suggest that PDS is a positive regulator of ripening in tomato fruit, which must be considered when using it as a marker for virus-induced gene silencing (VIGS) experiments in order to avoid fruit-ripening side effects.

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Characterization of Tomato Fruit LeRIN Gene Application Virus-Induced Gene Silencing Technology

Advanced Materials Research ◽

10.4028/www.scientific.net/amr.781-784.1685 ◽

2013 ◽

Vol 781-784 ◽

pp. 1685-1688

Author(s):

Ling Li ◽

Guo Xia Zhu ◽

Zheng Liu ◽

Tie Ling Liu

Keyword(s):

Gene Silencing ◽

Fruit Ripening ◽

Tomato Fruit ◽

Specific Gene ◽

Virus Induced Gene Silencing ◽

Green Color ◽

Tomato Fruit Ripening ◽

Infiltration Method ◽

I Gene

Virus-induced gene silencing (VIGS) is a powerful tool for the study of gene function and down regulate specific gene expression in plant. The RIN transcription factor is an important developmental regulator in tomato fruit ripening. In this work, the TRV-LeRIN infiltrated fruit significantly developed green color phenotypes, whereas the control fruit would turn red normally. Dissection of silenced areas for analysis identified whether the LeRIN gene is silenced or not. Taken together, we successfully applied syringe infiltration method of VIGS to silence the LeRIN gene in tomato fruit. These results are critical for understanding the mechanism of tomato fruit ripening.

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A comprehensive investigation of starch degradation process and identification of a transcriptional activator MabHLH6 during banana fruit ripening

Plant Biotechnology Journal ◽

10.1111/pbi.12756 ◽

2017 ◽

Vol 16 (1) ◽

pp. 151-164 ◽

Cited By ~ 38

Author(s):

Yun-yi Xiao ◽

Jian-fei Kuang ◽

Xin-na Qi ◽

Yu-jie Ye ◽

Zhen-Xian Wu ◽

...

Keyword(s):

Fruit Ripening ◽

Transcriptional Activator ◽

Degradation Process ◽

Starch Degradation ◽

Banana Fruit ◽

Comprehensive Investigation

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Virus-Induced Gene Silencing of the Eggplant Chalcone Synthase Gene during Fruit Ripening Modifies Epidermal Cells and Gravitropism

Journal of Agricultural and Food Chemistry ◽

10.1021/acs.jafc.7b05617 ◽

2018 ◽

Vol 66 (11) ◽

pp. 2623-2629 ◽

Cited By ~ 5

Author(s):

Cuicui Wang ◽

Daqi Fu

Keyword(s):

Gene Silencing ◽

Fruit Ripening ◽

Chalcone Synthase ◽

Epidermal Cells ◽

Virus Induced Gene Silencing ◽

Chalcone Synthase Gene

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Optimization of a virus-induced gene silencing system and functional elucidation of MaBAM9b in postharvest banana fruits

10.21203/rs.3.rs-31259/v2 ◽

2020 ◽

Author(s):

juhua liu ◽

Meng Li ◽

Jing Zhang ◽

Hongxia Miao ◽

Jingyi Wang ◽

...

Keyword(s):

Gene Silencing ◽

Reverse Genetics ◽

Starch Content ◽

Sugar Content ◽

Soluble Sugar ◽

Starch Degradation ◽

Virus Induced Gene Silencing ◽

Functional Identification ◽

Exact Function ◽

Starch Conversion

Abstract Background: The genetic basis of metabolic pathways that operate during fruit ripening needs to be understood before the nutritional value of the banana can be improved. The banana is a typical starch conversion fruit, and β-amylase is a key enzyme that may play an important role in starch degradation during the ripening process. Musa acuminata β-amylase 9b (MaBAM9b) is closely related to starch degradation. However, its exact function in starch degradation has not been demonstrated in banana. Stable genetic transformation to identify gene function is time- and energy-consuming. Thus, an efficient and rapid method is needed for functional identification. Virus-induced gene silencing (VIGS) is a reverse-genetics method based on RNA-mediated antiviral plant defense that has been used to rapidly identify gene functions in plants. The aim of this study is to optimize a VIGS system and functional elucidation of MaBAM9b in postharvest banana fruits. Results: Using 0.5% iodine-potassium-iodide (I2-KI) staining for 150 s, we found that 1:3 TRV1:TRV2-MaBAM9b cultivated at 30 mmHg for 30 s to an optical density (OD) of 0.8 at 600 nm, and kept on Murashige & Skoog (MS) media for 5 days produced the best silencing results. Under these conditions, the total starch content was greatly increased, whereas the β-amylase activity, soluble sugar content, and expression of endogenous MaBAM9b greatly decreased. Conclusions: The system described here is particularly useful for studying genes and networks involved in starch conversion in fruit, which alone would not produce a visual phenotype. This system will provide a platform for functional genomics and fruit quality improvement in the banana.

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