Efficient transgenic plant regeneration throughAgrobacterium-mediated transformation of Chickpea (Cicer arietinum L.)

Kar, Sanchayita; Johnson, Tony M.; Nayak, Pritilata; Sen, Sandeep

doi:10.1007/bf01275444

Cited by 80 publications

(11 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Direct shoot organogenesis and establishment of plantlets from different explants of chickpea was reported earlier (Polisetty et al, 1996(Polisetty et al, , 1997Paul et al, 2000;Rizvi and Singh, 2000;Chauhan et al, 2003;Jayanand et al, 2003;Chakraborti et al, 2006). Plantlets were developed through direct somatic embryogenesis and through callus from different explants of chickpea (Barna and Wakhlu, 1993;Sagare et al, 1993;Suhasini et al, 1994;Kumar et al, 1994Kumar et al, , 1995Rizvi and Singh, 2000;Chauhan et al, 2002;Kar et al, 1996Kar et al, , 1997Kiran et al, 2005).…”

Section: Introductionmentioning

confidence: 74%

An optimal protocol for in vitro regeneration, efficient rooting and stable transplantation of chickpea

Anwar

Sharmila

Saradhi

2008

Physiol Mol Biol Plants

View full text Add to dashboard Cite

A rapid, reproducible and efficient regeneration method was developed for chickpea (Cicer arietinum L.) using single cotyledon with half embryonal axis as explants. MS medium supplemented with 4 ìM TDZ, 10 ìM 2-iP and 2 ìM kinetin induced 50-100 adventitious buds/shoots after 14 days of culture and elongated on MS medium supplemented with 5 ìM 2-iP and 2 ìM kinetin. Healthy, strong and 100 % rooting was achieved by exposing cut ends of the shoots to 10 sec pulse treatment with 100 ìmoles/ml IBA followed by their transfer to liquid MS basal medium within 10-14 d. 2-3 cm long shoots were most suitable for rooting. Potting-mixture with good aeration and lesser capacity to retain water was most suitable for achieving successful establishment of chickpea plantlets. Garden soil mixed with sand (gravel) and bio-manure in the ratio of 1:1:1 is most suitable for achieving cent percent transplantation success. Cent percent of plantlets got acclimatized, survived in the pots and showed normal growth, development, flowering followed by podding and seeds setting. Harvesting of seeds was done after the pods were fully matured and dry. In this communication, we have demonstrated for the first time that shoot length, pulse treatment of cut ends of shoots with 100 ìmoles/ml IBA and aeration of potting mixture are key factors for rapid micro-propagation and successful establishment of chickpea.

show abstract

Section: Introductionmentioning

confidence: 74%

An optimal protocol for in vitro regeneration, efficient rooting and stable transplantation of chickpea

Anwar

Sharmila

Saradhi

2008

Physiol Mol Biol Plants

View full text Add to dashboard Cite

show abstract

“…Three-to 4-week-old calli and excised embryo-axes from 3 to 4-d-old germinated immature seeds exhibited great regeneration potential, which subsequently declines with increasing age of the target cells. We have clearly demonstrated that tissues which respond well in tissue culture, also tended to respond well In most of the previous reports on Agrobacteriummediated transformation of chickpea, either embryo axes or slices of embryo axes were used as the target tissue (Fontana et al 1993, Kar et al 1996, Krishnamurthy et al 2000, Polowick et al 2004, Senthil et al 2004) and two of the recent reports were based on the decapitated embryo explant attached with one half of the cotyledon (Sarmah et al 2004, Tewari-Singh et al 2004. However, in the present study, one-month-old, dark-green cotyledonary-node-derived calli showed the highest transformation efficiency and was found to be the best source of explant for regeneration of complete plant from all the four genotypes compared to any other source of explants used.…”

Section: Discussionmentioning

confidence: 94%

“…For example, Fontana et al (1993) claimed a 4 % success rate to achieve only 3 GUS positive transgenic chickpea plants. Kar et al (1996), Krishnamurthy et al (2000) and Tewari-Singh et al (2004) reported Agrobacterium-mediated transformation of chickpea based on multiple shoot formation from embryo axis explants. Only Krishnamurthy et al (2000) and Tewari-Singh et al (2004) reported the inheritance of transgene in T 1 generation of transformed plants of chickpea.…”

Section: Introductionmentioning

confidence: 98%

Transformation of chickpea: effect of genotype, explant, Agrobacterium-strain and composition of culture medium

Bhattacharjee¹,

Mohan²,

Nair³

2010

Biologia plant.

View full text Add to dashboard Cite

Reproducible and high-frequency transgenic plant regeneration from callus and embryo axes of four different genotypes of chickpea (Cicer arietinum) was achieved after Agrobacterium-mediated transformation. Three different strains of Agrobacterium (EHA105, AGL1 and LBA4404) harboring the binary vector pCAMBIA1301 containing β-glucuronidase (GUS) and hygromycin phosphotransferase (hpt) genes under the control of a CaMV35S promoter were used. The highest number of transgenic plants was obtained from cotyledonary node-derived calli of genotype Pusa-256. A highly efficient rooting was achieved on Murashige and Skoog medium supplemented with indole-3-butyric acid. The stable integration of the gene was confirmed by molecular analyses of the transformed plants. Inheritance of GUS and hpt gene was followed through two generations and they showed the expected 3:1 inheritance.Additional key words: Cicer arietinum, cotyledon-and cotyledonary node-derived calli, β-glucuronidase, hygromycin phosphotransferase.

show abstract

“…Furthermore, there are limited reports of reliable rooting and establishment of in vitro regenerated shoots in soil. Probably due to these difficulties the genetic transformation of chickpea (Fontana et al 1993;Kar et al 1996;Polowick et al 2004;Senthil et al 2004) has not made any significant progress as of now. The few reports so far published by independent research groups on transgenic chickpea plants for developing resistance against either pod borer or storage pests, were using particle bombardment or Agrobacterium-mediated transformation technique (Kar et al 1997;Sarmah et al 2004;Sanyal et al 2005;Indurker et al 2007).…”

Section: Discussionmentioning

confidence: 97%

Tissue specific expression of potent insecticidal, Allium sativum leaf agglutinin (ASAL) in important pulse crop, chickpea (Cicer arietinum L.) to resist the phloem feeding Aphis craccivora

et al. 2009

View full text Add to dashboard Cite

The phloem sap-sucking hemipteran insect, Aphis craccivora, commonly known as cowpea aphid, cause major yield loss of important food legume crop chickpea. Among different plant lectins Allium sativum leaf agglutinin (ASAL), a mannose binding lectin was found to be potent antifeedant for sap sucking insect A. craccivora. Present study describes expression of ASAL in chickpea through Agrobacterium-mediated transformation of "single cotyledon with half embryo" explant. ASAL was expressed under the control of CaMV35S promoter for constitutive expression and phloem specific rolC promoter for specifically targeting the toxin at feeding site, using pCAMBIA2301 vector containing plant selection marker nptII. Southern blot analysis demonstrated the integration and copy number of chimeric ASAL gene in chickpea and its inheritance in T(1) and T(2) progeny plants. Expression of ASAL in T(0) and T(1) plants was confirmed through northern and western blot analysis. The segregation pattern of ASAL transgene was observed in T(1) progenies, which followed the 3:1 Mendelian ratio. Enzyme linked immunosorbant assay (ELISA) determined the level of ASAL expression in different transgenic lines in the range of 0.08-0.38% of total soluble protein. The phloem tissue specific expression of ASAL gene driven by rolC promoter has been monitored by immunolocalization analysis of mature stem sections. Survival and fecundity of A. craccivora decreased to 11-26% and 22-42%, respectively when in planta bioassay conducted on T(1) plants compared to untransformed control plant which showed 85% survival. Thus, through unique approach of phloem specific expression of novel insecticidal lectin (ASAL), aphid resistance has been successfully achieved in chickpea.

show abstract

Efficient transgenic plant regeneration throughAgrobacterium-mediated transformation of Chickpea (Cicer arietinum L.)

Cited by 80 publications

References 24 publications

An optimal protocol for in vitro regeneration, efficient rooting and stable transplantation of chickpea

An optimal protocol for in vitro regeneration, efficient rooting and stable transplantation of chickpea

Transformation of chickpea: effect of genotype, explant, Agrobacterium-strain and composition of culture medium

Tissue specific expression of potent insecticidal, Allium sativum leaf agglutinin (ASAL) in important pulse crop, chickpea (Cicer arietinum L.) to resist the phloem feeding Aphis craccivora

Contact Info

Product

Resources

About