Genetic Transformation of Plumbago zeylanica with Agrobacterium rhizogenes Strain LBA 9402 and Characterization of Transformed Root Lines

Basu, Amrita; Joshi, Raj Kumar; Jha, Sumita

doi:10.3329/ptcb.v25i1.24123

Cited by 16 publications

(4 citation statements)

References 35 publications

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“…The CDSs of BrrICE1.1 and BrrADC2.2 and their reverse complementary sequences were cloned into PRI101-AN-Flag vectors. Then, these constructs were separately transferred into A. rhizogenes LBA9402 by electroporation [ 47 ]. Cotyledon infection was used to transform the above genes into turnip roots, and the gene expression levels and putrescine content in the hair roots were extracted and analyzed by qRT-PCR and LC-MS, respectively, as described above.…”

Section: Methodsmentioning

confidence: 99%

BrrICE1.1 is associated with putrescine synthesis through regulation of the arginine decarboxylase gene in freezing tolerance of turnip (Brassica rapa var. rapa)

Yin

Yang

et al. 2020

BMC Plant Biol

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Background In the agricultural areas of Qinghai-Tibet Plateau, temperature varies widely from day to night during the growing season, which makes the extreme temperature become one of the limiting factors of crop yield. Turnip (Brassica rapa var. rapa) is a traditional crop of Tibet grown in the Tibet Plateau, but its molecular and metabolic mechanisms of freezing tolerance are unclear. Results Here, based on the changes in transcriptional and metabolic levels of Tibetan turnip under freezing treatment, the expression of the arginine decarboxylase gene BrrADC2.2 exhibited an accumulative pattern in accordance with putrescine content. Moreover, we demonstrated that BrrICE1.1 (Inducer of CBF Expression 1) could directly bind to the BrrADC2.2 promoter, activating BrrADC2.2 to promote the accumulation of putrescine, which was verified by RNAi and overexpression analyses for both BrrADC2.2 and BrrICE1.1 using transgenic hair root. The function of putrescine in turnip was further analyzed by exogenous application putrescine and its inhibitor DL-α-(Difluoromethyl) arginine (DFMA) under freezing tolerance. In addition, the BrrICE1.1 was found to be involved in the ICE1-CBF pathway to increase the freezing stress of turnip. Conclusions BrrICE1.1 could bind the promoter of BrrADC2.2 or CBFs to participate in freezing tolerance of turnip by transcriptomics and targeted metabolomics analyses. This study revealed the regulatory network of the freezing tolerance process in turnip and increased our understanding of the plateau crops response to extreme environments in Tibet.

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Section: Methodsmentioning

confidence: 99%

BrrICE1.1 is associated with putrescine synthesis through regulation of the arginine decarboxylase gene in freezing tolerance of turnip (Brassica rapa var. rapa)

Yin

Yang

et al. 2020

BMC Plant Biol

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“…This is due to the high density of bacteria that can damage cells or plant tissue. Basu et al (2015) study on Plumbago zeilanica reported that the OD 600 ˃ 0.1 is best density of Agobacterium rhizogenes strain LBA 9402.…”

Section: Resultsmentioning

confidence: 99%

Hairy Root Induction on Justicia gendarussa by Various Density of Agrobacterium rhizogenes strain LB 510

et al. 2017

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Gandarusa (Justicia gendarussa Burm.f.) is an Indonesian medicinal plant that has many benefits as drug and male contracetive. For industrial needs, Gandarusa must be available in large quantity. Hairy root culture is one of methode to produce phytochemistry compound. The objective of the study was to examine the effect of various density of Agrobacterium rhizogenes strain LB510 on hairy roots induction of gandarusa (Justicia gendarussa Burm.f.) leaf plant. Leaf explants were inoculated in MS liquid medium with various density of OD 600 = 0.1; 0.2; 0.3; 0.4; and 0.5. Explants were co-cultivated for 2 days on MS solid medium without any hormone then sub-cultured on MS solid medium containing antibiotic cefotaxim 300 ppm, in dark condition. The data were analyzed descriptively and statistically. The results showed that various density of Agrobacterium rhizogenes strain LB510 was affected the lenght of hairy roots induction of J. gendarussa Burm.f., but these was not effected toward lenght formation time and number of hairy root. The treatment of OD 600 0.2 was the best treatment for hairy root induction on Justicia gendarussa Burm. f. This data could be used for optimized the quality of methode of hairy root induction. preliminary study is needed to determine the appropriate method for the hairy roots induction of gandarusa. Therefore, this study was aimed to obtain scientific information on the effect of the Agrobacterium rhizogenes strain LB510 density toward gandarusa hairyroots induction. How to CiteA protocol for hairy root induction of Justicia gendarussa Burm.f. has not been established and the present study of the influence of some factor, expecially Agrobacterium rhizogenes density, on hairy root induction in Justicia gendarussa Burm. f. was an attempt to develop a technique for hairy root production in Justicia gendarussa Burm.f. METHODS Plant preparationPlants used as explants are leaves of gandarusa (Justicia gendarussa Burm.f.). This plant was obtained from the Institute of Materia Medical, Batu, Malang, Indonesia. Leaves were taken from the second-third nodal segment. The leaves were washed under running water for ten minute and then disinfected by anti-fungiside "topsin" with 1% (v/v), followed by several rinses in sterilized distilled water and sterilized by 10% Clorox "baycline" for ten minute, followed by three times rinses in sterilized distilled water.Bacterial strain and culture media LB510 strain of Agrobacterium rhizogenes was obtained from the Research Center of Biotechnology, Indonesian Institute of Sciences (LIPI), Bogor, Indonesia. The LB510 strain was grown at room temperature (±27°C) in Luria Bertani medium (LB) (per l) (10g trypton, 5g yeast extract and 10g NaCl) of solid and liquid. Media was sterilized by autoclave at 1.2 atm 121°C for 20 minutes.The bacteria was stroke on solid media. Isolates were stored in the refrigerator for 1 week. The bacteria has been rejuvenated to each 25 mL of YMB medium and liquid LB medium. Then left at room temperature for 24 hours. Agrobacteri...

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“…There are contradictory reports regarding the extent of stability in transformed roots maintained in vitro during prolonged culture. Stability in growth and morphological characters over prolonged culture is reported in different species like Lycopersicon esculentum [518], Cinchona officinalis "Ledgeriana" [519], Aesculus hippocastanum [520], Pimpinella anisum [521], Plumbago zeylanica [522], Rauvolfia serpentina [522], Tylophora indica [523], Bacopa monnieri [524], etc. However, instability in morphological phenotype and growth kinetics of hairy roots in prolonged in vitro culture is reported in certain species like carrot, Brugmansia candida, Duboisia myoporoides, Hyoscyamus muticus, etc.…”

Section: Hairy Root Culturesmentioning

confidence: 99%

Morphogenesis, Genetic Stability, and Secondary Metabolite Production in Untransformed and Transformed Cultures

Halder¹,

Jha²

2020

Reference Series in Phytochemistry

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In addition to the primary metabolites, plants produce a vast number of chemically diversified economically valuable secondary metabolites (SMs) in specific cell types or tissue or organ of phylogenetically related family or genus or species or even to a single chemical race under tight environmental, developmental, and genetic control. Some SMs such as morphine and codeine are only synthesized in a particular organized and differentiated tissue or organ.

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Genetic Transformation of Plumbago zeylanica with Agrobacterium rhizogenes Strain LBA 9402 and Characterization of Transformed Root Lines

Cited by 16 publications

References 35 publications

BrrICE1.1 is associated with putrescine synthesis through regulation of the arginine decarboxylase gene in freezing tolerance of turnip (Brassica rapa var. rapa)

BrrICE1.1 is associated with putrescine synthesis through regulation of the arginine decarboxylase gene in freezing tolerance of turnip (Brassica rapa var. rapa)

Hairy Root Induction on Justicia gendarussa by Various Density of Agrobacterium rhizogenes strain LB 510

Morphogenesis, Genetic Stability, and Secondary Metabolite Production in Untransformed and Transformed Cultures

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