rol genes of Agrobacterium rhizogenes cucumopine strain: sequence, effects and pattern of expression

Serino, Giovanna; Clérot, Danielle; Brevet, Jean; Costantino, Paolo; Cardarelli, Maura

doi:10.1007/bf00039550

Cited by 32 publications

(15 citation statements)

References 34 publications

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“…The sequences used for the alignment were collected from the PIR protein sequence database. Accession numbers are as follows: 1724RolB, (Tanaka et al ., 1994); 2659RolB, (Serino et al ., 1994); A4RolB, (Furner et al ., 1986); 8196RolB, (Hansen et al ., 1991); NgRolB, (Furner et al ., 1986). (c) Mutations of amino acid residues of 1724RolB protein that impair interaction with Nt14‐3‐3 ωII.…”

Section: Resultsmentioning

confidence: 99%

Nuclear localization and interaction of RolB with plant 14‐3‐3 proteins correlates with induction of adventitious roots by the oncogene rolB

et al. 2004

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SummaryThe rooting-locus gene B (rolB) on the T-DNA of the root-inducing (Ri) plasmid in Agrobacterium rhizogenes is responsible for the induction of transformed adventitious roots, although the root induction mechanism is unknown. We report here that the RolB protein of pRi1724 (1724RolB) is associated with Nicotiana tabacum 14-3-3-like protein vII (Nt14-3-3 vII) in tobacco bright yellow (BY)-2 cells. Nt14-3-3 vII directly interacts with 1724RolB protein. Green¯uorescent protein (GFP)-fused 1724RolB is localized to the nucleus. GFP-fused mutant 1724RolB proteins having a deletion or amino acid substitution are unable to interact with Nt14-3-3 vII and also show impaired nuclear localization. Moreover, these 1724RolB mutants show decreased capacity for adventitious root induction. These results suggest that adventitious root induction by 1724RolB protein correlates with its interaction with Nt14-3-3 vII and the nuclear localization of 1724RolB protein.

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Section: Resultsmentioning

confidence: 99%

Nuclear localization and interaction of RolB with plant 14‐3‐3 proteins correlates with induction of adventitious roots by the oncogene rolB

et al. 2004

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“…This may be due to the reason that hairy roots were induced by the cucumopine-type A. rhizogenes K599, whose T-DNA contained only 11 genes but none of them encode the proteins involved in the biosynthesis of any plant hormones (Mankin et al 2007). This is different from hairy roots induced by infection with the agropine type A. rhizogenes (such as A4 strain) (Serino et al 1994).…”

Section: Discussionmentioning

confidence: 99%

Transgenic hairy roots of Tetrastigma hemsleyanum: induction, propagation, genetic characteristics and medicinal components

Song

et al. 2015

Plant Cell Tiss Organ Cult

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Tetrastigma hemsleyanum Diels & Gilg ex Diels (T. hemsleyanum) is an important medicinal plant in China. Its bulbous root is the most valuable part for medicine. There is an increasing demand for T. hemsleyanum in the medicinal market. However, the natural slow growth rate of T. hemsleyanum hardly meets the demand of the medicinal market. There is the need of alternative way for rapid and large-scale production of T. hemsleyanum. In this study, we investigated the induction of hairy roots by using Agrobacterium rhizogenes (A. rhizogenes) as an alternative method of increasing production of the secondary metabolites in hairy roots of T. hemsleyanum. Herein, we reported our successful induction of hairy roots of T. hemsleyanum by cucumopine type A. rhizogenes K599 and their rapid propagation. Firstly, we selected the optimal mediums for induction of hairy roots and for subculture of induced hairy roots. We found that when A. rhizogenes K599 carrying both an endogenous plasmid pRi2659 and an exogenous plasmid pRI101-AN-GFP was used to infect the leaves of T. hemsleyanum seedlings, the optimal medium for inducting hairy roots was the medium of MS ? 0.4 mg/L 1-naphthaleneacetic acid. The optimal medium for subculture of hairy roots was the combination of MS ? 1.0 mg/L indole-3-butyric acid ? 0.5 or 1.0 mg/L kinetin. During suspension culture of hairy roots, the rapid growth phase occurred in the period of 15-28 days. Secondly, we employed polymerase chain reaction (PCR) assays to examine the integration of T-DNA of A. rhizogenes. The PCR results revealed the existence of a phenomenon of lacking of integration of one or more of 11 genes within T-DNA of plasmid pRi2659 into the genomic DNA of hairy roots. Thirdly, we conducted histocytological analysis to examine the structural features of the induced hairy roots and observed that xylem structures of the hairy roots were simplified as compared to those of the bulbous roots and ordinary fine roots of non-transgenic plant. Finally, we applied high performance liquid chromatography to analyze the medicinal components in several organs of the T. hemsleyanum plants. The results showed that the contents of kaempferol in all hairy root lines were significantly higher than those in bulbous roots, fine roots, stems and leaves. These results suggest that insertion and integration of T-DNA from A. rhizogenes can significantly increase the kaempferol content in hairy roots of T. hemsleyanum.

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“…Sequences of PCR primers used for PCR‐restriction fragment length polymorphism (RFLP) in this study are listed in Table 2. Four primer sets orf3+4 , orf8+10 , orf13 and cus were designed to give products of around 1500 bp using Primer Express 2.0 software (Applied Biosystems, Foster City, CA) from the published 16110 bp T‐DNA sequence (Serino et al , 1994; GenBank Accession no. ) of a known RMA Agrobacterium radiobacter (NCPPB 2659) pRi.…”

Section: Methodsmentioning

confidence: 99%

Recurrent outbreaks of root mat in cucumber and tomato are associated with a monomorphic, cucumopine, Ri-plasmid harboured by various Alphaproteobacteria

Weller¹,

Stead²,

Young³

2006

FEMS Microbiology Letters

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Recurrent outbreaks of root mat have occurred in the UK and France in cucumber and tomato. Root mat is caused by bacterial strains harbouring a Ri-plasmid (pRi). Fifteen root mat-associated (RMA) cucumopine pRi were analysed by PCR-restriction fragment length polymorphism (RFLP) and Southern blotting. These pRi were harboured by Agrobacterium biovar 1 strains isolated during a 1970s outbreak of root mat in UK soil grown cucumber, and also by Agrobacterium biovar 1, Ochrobactrum, Rhizobium and Sinorhizobium isolated during outbreaks of root mat in cucumber and tomato grown hydroponically in the UK and France since 1993. PCR-RFLP analysis of the T-DNA and virD2 regions showed sequence homology between all cucumopine pRi, indicating that these pRi are monomorphic, and thus this pRi remained in the UK without inducing symptoms for some 15 years between outbreaks in the 1970s and 1990s. Cucumopine pRi were also shown to possess the virE2 substitute GALLS gene by Southern blotting. Two other pRi, harboured by Agrobacterium isolated from a recent root mat outbreak in one tomato crop, were also shown to possess the GALLS gene but were shown not to be cucumopine pRi by PCR-RFLP.

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rol genes of Agrobacterium rhizogenes cucumopine strain: sequence, effects and pattern of expression

Cited by 32 publications

References 34 publications

Nuclear localization and interaction of RolB with plant 14‐3‐3 proteins correlates with induction of adventitious roots by the oncogene rolB

Nuclear localization and interaction of RolB with plant 14‐3‐3 proteins correlates with induction of adventitious roots by the oncogene rolB

Transgenic hairy roots of Tetrastigma hemsleyanum: induction, propagation, genetic characteristics and medicinal components

Recurrent outbreaks of root mat in cucumber and tomato are associated with a monomorphic, cucumopine, Ri-plasmid harboured by various Alphaproteobacteria

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