Agrobacterium-mediated Transformation in Chickpea (Cicer Arietinum L.) with an Insecticidal Protein Gene: Optimisation of Different Factors

Overview

Journal Physiol Mol Biol Plants

Specialty Biology

Date 2013 Apr 11

PMID 23572977

Citations 12

Authors

Shivani Indurker

Hari S Misra

Susan Eapen

Affiliations

Soon will be listed here.

Abstract

Agrobacterium-mediated transformation in chickpea was developed using strain LBA4404 carrying nptII, uidA and cryIAc genes and transformants selected on Murashige and Skoog's basal medium supplemented with benzyladenine, kinetin and kanamycin. Integration of transgenes was demonstrated using polymerase chain reaction and Southern blot hybridization of T0 plants. The expression of CryIAc delta endotoxin and GUS enzyme was shown by enzyme linked immunosorbent assay and histochemical assay respectively. The transgenic plants (T0) showed more tolerance to infection by Helicoverpa armigera compared to control plants. Various factors such as explant source, cultivar type, different preculture treatment period of explants, co-cultivation period, acetosyringone supplementation, Agrobacterium harboring different plasmids, vacuum infiltration and sonication treatment were tested to study the influence on transformation frequency. The results indicated that use of epicotyl as explant, cultivar ICCC37, Agrobacterium harboring plasmid pHS102 as vector, preculture of explant for 48 h, co-cultivation period of 2 days at 25°C and vacuum infiltration for 15 min produced the best transformation results. Sonication treatment of explants with Agrobacteria for 80 s was found to increase the frequency of transformation.

Citing Articles

Recalcitrance to transformation, a hindrance for genome editing of legumes.

Nivya V, Shah J Front Genome Ed. 2023; 5:1247815.

PMID: 37810593 PMC: 10551638. DOI: 10.3389/fgeed.2023.1247815.

Unclasping potentials of genomics and gene editing in chickpea to fight climate change and global hunger threat.

Singh C, Kumar R, Sehgal H, Bhati S, Singhal T, Gayacharan Front Genet. 2023; 14:1085024.

PMID: 37144131 PMC: 10153629. DOI: 10.3389/fgene.2023.1085024.

Activity of Arabidopsis Rubisco small subunit promoter in various tissues of chickpea.

Boruah R, Konwar T, Nath P, Acharjee S, Sarmah B 3 Biotech. 2023; 13(3):89.

PMID: 36815010 PMC: 9939566. DOI: 10.1007/s13205-023-03508-z.

Chickpea ( L.) Biology and Biotechnology: From Domestication to Biofortification and Biopharming.

Koul B, Sharma K, Sehgal V, Yadav D, Mishra M, Bharadwaj C Plants (Basel). 2022; 11(21).

PMID: 36365379 PMC: 9654780. DOI: 10.3390/plants11212926.

Mediated Genetic Transformation of Taiwanese Isolates of .

Wang K, Hong M, Wu Y, Wu T Plants (Basel). 2021; 10(8).

PMID: 34451621 PMC: 8401387. DOI: 10.3390/plants10081576.

References

Park B, Liu Z, Kanno A, Kameya T . Transformation of radish (Raphanus sativus L.) via sonication and vacuum infiltration of germinated seeds with Agrobacterium harboring a group 3 LEA gene from B. napus. Plant Cell Rep. 2005; 24(8):494-500. DOI: 10.1007/s00299-005-0973-5. View

Paz M, Martinez J, Kalvig A, Fonger T, Wang K . Improved cotyledonary node method using an alternative explant derived from mature seed for efficient Agrobacterium-mediated soybean transformation. Plant Cell Rep. 2005; 25(3):206-13. DOI: 10.1007/s00299-005-0048-7. View

Tewari-Singh N, Sen J, Kiesecker H, Reddy V, Jacobsen H, Guha-Mukherjee S . Use of a herbicide or lysine plus threonine for non-antibiotic selection of transgenic chickpea. Plant Cell Rep. 2004; 22(8):576-83. DOI: 10.1007/s00299-003-0730-6. View

Liu H, Yang C, Wei Z . Efficient Agrobacterium tumefaciens-mediated transformation of soybeans using an embryonic tip regeneration system. Planta. 2004; 219(6):1042-9. DOI: 10.1007/s00425-004-1310-x. View

Jaiwal P, Kumari R, Ignacimuthu S, Potrykus I, Sautter C . Agrobacterium tumefaciens-mediated genetic transformation of mungbean (Vigna radiata L. Wilczek) - a recalcitrant grain legume. Plant Sci. 2001; 161(2):239-247. DOI: 10.1016/s0168-9452(01)00352-1. View

Ke J, Khan R, Johnson T, Somers D, Das A . High-efficiency gene transfer to recalcitrant plants by Agrobacterium tumefaciens. Plant Cell Rep. 2019; 20(2):150-156. DOI: 10.1007/s002990000271. View

Kar S, Johnson T, Nayak P, Sen S . Efficient transgenic plant regeneration throughAgrobacterium-mediated transformation of Chickpea (Cicer arietinum L.). Plant Cell Rep. 2013; 16(1-2):32-7. DOI: 10.1007/BF01275444. View

Polowick P, Baliski D, Mahon J . Agrobacterium tumefaciens-mediated transformation of chickpea (Cicer arietinum L.): gene integration, expression and inheritance. Plant Cell Rep. 2004; 23(7):485-91. DOI: 10.1007/s00299-004-0857-0. View

Srinivasan , Mohapatra T, Sharma R . Agrobacterium mediated genetic transformation of chickpea, Cicer arietinum L. Indian J Exp Biol. 1991; 29(8):758-61. View

10.

Davies D, Hamilton J, Mullineaux P . Transformation of peas. Plant Cell Rep. 2013; 12(3):180-3. DOI: 10.1007/BF00239102. View

11.

Trick H, Finer J . Sonication-assisted Agrobacterium-mediated transformation of soybean [Glycine max (L.) Merrill] embryogenic suspension culture tissue. Plant Cell Rep. 2019; 17(6-7):482-488. DOI: 10.1007/s002990050429. View

12.

Santarem E, Trick H, Essig J, Finer J . Sonication-assisted Agrobacterium-mediated transformation of soybean immature cotyledons: optimization of transient expression. Plant Cell Rep. 2019; 17(10):752-759. DOI: 10.1007/s002990050478. View

13.

Fontana G, Santini L, Caretto S, Frugis G, Mariotti D . Genetic transformation in the grain legume Cicer arietinum L. (chickpea). Plant Cell Rep. 2013; 12(4):194-8. DOI: 10.1007/BF00237052. View

14.

Senthil G, Williamson B, Dinkins R, Ramsay G . An efficient transformation system for chickpea (Cicer arietinum L.). Plant Cell Rep. 2004; 23(5):297-303. DOI: 10.1007/s00299-004-0854-3. View

15.

Krishnamurthy K, Suhasini K, Sagare A, Meixner M, de Kathen A, Pickardt T . Agrobacterium mediated transformation of chickpea (Cicer arietinum L.) embryo axes. Plant Cell Rep. 2019; 19(3):235-240. DOI: 10.1007/s002990050005. View