Table 1.

Transformation of Sorghum via Agrobacterium and Biolistic Approach.

Explants	Agrobacterium Strain	Vectors	Selection Marker	Promoter Used for Selection Marker	Reporter Gene	Promoter Used for Reporter Gene	Transgene Detection/Expression	Transformation Efficiency	Key Points/Studied Parameters	References
Immature Embryo	LBA4404	pSB1 pSB11	bar	Ubi1	gus	Ubi1	Southern Blot	2.1%	Study on protocol optimization and source of genotype.	Zhao et al. [19]
Immature Embryo	EHA105 EHA101 AGL1	pPZP200	hbt	Ubi1, CaMV35S	gfp, gus	Hbt chimeric promoter with CaMV35S enhancer	PCR	Only a few events	Compare gfp and gus reporter genes by direct and indirect gene transfer methods.	Jeoung et al. [13]
Immature Embryo	LBA4404	pTOK233	hpt	CaMV35S	gus	CaMV35S	PCR, Southern Blot	1.7–3.5%	Media optimization by antioxidants and study transient gus expression.	Carvelho et al. [10]
Immature Embryo	EHA101	pPZP201	pmi	Ubi1	gfp	Ub1	Southern Blot, Western Blot, CPR Assay, PCR	2.8–3.3%	Study pmi gene as a selectable marker on mannose selective agent.	Gao et al. [20]
Immature Embryo	NTL4	pPZP212	npt II	Ubi1	gus	Ubi1	Southern blot	0.3–4.5%	Utilized standard binary vector with npt II gene as a selectable marker.	Howe et al. [22]
Immature Embryo	LBA4404	pCAMBIA1301	hpt	CaMV35S	gus	CaMV35S	Southern Blot PCR	5%	Apply cold-pre-treatment on explant, increase callus induction and reduced phenols.	Nguyen et al. [25]
Immature Embryo	EHA101 LBA4404	pPZP201	pmi	Ubi1	gfp	Ubi1	PCR, Western Blot	8.3%	Treatment to immature embryos at 43°C, with various time frames.	Gurel et al. [6]
Immature Inflorescence	EHA105	pKUB	hpt	Ubi1	gus	CaMV35S	RT-PCR, Southern Blot, Western Blot	1.9%	Incorporation of insecticidal cry1Ab gene.	Zhang et al. [58]
Immature Embryo	EHA105, EHA101, LBA4404	pMKURF2, pCAMBIAG11 pCAMBIARC7	hpt	Ubi1	gus	Ubi1	Western blot	1.6–2.7%	Study of pathogen resistance, chitinase.	Arulselvi et al. [59]
Mature Embryo, Young Seedling, Immature Inflorescence	LBA4404	pKU352NA	hpt	Ubi1	SgfpS65T,	Ubi1	Inverse PCR	4.28%	Improved gfp and Ac-Ds system.	Jambagi et al. [23]
Immature Embryo	NTL4	pCAMBIA1305.2 pCAM-UBIgus	hpt	Ubi1	gus	Ubi1	PCR, Southern Blot	2.4%	Study the effect of L-cystine and inclusion of additional binary vector.	Kumar et al. [38]
Shoot Apices	EHA105	pCAMBIA1305			gus	CaMV35S	Histochemical gus assay	Few transient event	Effect of cysteine on gus activity.	Pandey et al. [41]
Shoot Apices	LBA4404	pCAMBIA1305.1	hpt	CaMV35S	gus	CaMV35S	PCR, Southern Blot	1.2–3.9%	Insect resistance, cry1gene	Ignacimuthu and Premkumar [60]
Immature Embryo	LBA4404 AGL1	pSB1 pSB11	pmi	Ubi1	DsRed	Ubi1	QPCR	10% by LBA4404 & 33% by AGL1	Effect of CuSO4 and BAP in resting and selection media.	Wu et al. [40]
Immature Embryo	AGL1 EHA101 GV3101	pZY102, pFGC5941 pFGC161	bar	MAS, ZmUbi1	gus	CaMV35S	PCR Southern blot	14%	Study standard binary vector and bar gene as a selectable marker.	Do et al. [39]
Mature Embryo	EHA105	pCAMBIA1305.1 pCUbi1390	hph	Ubi1	gfp	CaMV35S	PCR, Western Blot	4%	Piercing the mature seeds excluding the tissue culture process	Li et al. [50]
Immature Embryo	EHA105	pHP78891		Ubi1	gfp	Ubi1	PCR, Southern Blot	6.2%	Study of morphogenic regulator, BABY BOOM, WUSCHEL2	Mookkan et al. [61]
Gene Transformation Through Biolistic Approach
Immature Embryo	biolistic	pBCI pNGI	hpt	Adh1	gus	Adh1	Gus Assay, RNA gel blot analysis	Observe transient events	Study of hygromycine and kanamycine resistance gene.	Hagio et al. 1991 [62]
Immature Embryo	biolistic	pPHP620 pPHP687	bar	D-CaMV35S	gus	D- CaMV35S	Southern Blot Gus assay	0.08%	Introduction of the bar gene.	Casas et al. 1993 [37]
Immature Embryo	biolistic		bar				Southern Blot, Western Blot, PCR	0.09%	Introduction of chitinase G11 gene.	Zhu et al. [8]
Immature Embryo, Leaf Segment	biolistic	pAHC20	bar	Ubi1, Actin, CaMV35S	gfp	Ubi1 Actin CaMV35S	Southern Blot	1%	Comparison of promoters and optimizing of PIG parameters.	Able et al. 2001 [63]
Immature Embryo	biolistic	pAHC20 pAct1-D	bar	Ubi1	gus	Actin	Southern Blot	0.18%	Methylation based Silencing of Act1-D	Emani et al. 2002 [64]
Immature Embryo, Mature Embryo, Shoot Tips	biolistic	pAct1-D pAHC25	bar, neo, hpt	Ubi1, Adh1, CaMV35S, ActD	gus	Ubi1, Adh1 CaMV35S Act1D	Southern Blot, Gus Assay, PCR	Few events	Tested physical parameters along with different promoters	Tadesse et al. [65]
Immature Embryo	biolistic	pPH1687	hpt, npt II	Ubi1	luc	Ubi1	Southern Blot	0.09%	Optimizing tissue culture parameters	Raghuwanshi and Birch [66]
Immature Embryo	biolistic	pAHC25 pNOV3604	bar, pmi	Ubi1	gus	Ubi1	PCR, Southern Blot	0.77%	Study the bar and pmi as selectable marker efficiency.	Grootboom et al. 2010 [67]
Immature Embryo	biolistic	pUKN pGEM-Ubi-gfp	npt II	Ubi1	gfp	Ubi1	PCR, Southern Blot	20.7%	Study the impact of Co bombardment of npt II and gfp, CuSO4	Liu and Godwin [68]

bar, Bialaphos resistance; CaMV35S, Cauliflower mosaic virus; gfp, Green fluorescent protein; gus, β-glucoronidase; hpt, hph, Hygromycin phosphotransferase; luc⁺, Luciferase; MAS, Mannopine synthase; npt, Neomycin phosphotransferase; pmi, Phosphomannose isomerase; Zm-Ubi1, Maize Ubiquitin1.