. Author manuscript; available in PMC: 2023 Sep 12.

Published in final edited form as: Nat Protoc. 2019 Sep 18;14(10):2954–2971. doi: 10.1038/s41596-019-0208-9

Table 1 |.

Summary and comparison of the DNA delivery methods in intact plants

Delivery platform		Target range	Genetic fate of target	Advantages	Platform limitations	Time line leading up to delivery to intact plants
Agrobacterium-mediated delivery		Mature plants, immature tissue (calli, meristems, embryos), protoplasts	Gene integration into genome, transient or stable expression, transformation of germ-line cells	Amenable to large sizes of DNA, high efficiency, stable transformation, no specialized equipment needed	High host specificity, time-consuming protocol, pathogenic and therefore regulated as GMO	Days 1–3: Agrobacterium-competent cell preparation Days 4 and 5: Agrobacterium transformation Days 6 and 7: preparation of Agrobacterium inoculum Day 8: Agrobacterium activation and infiltration
Biolistic delivery		Mature plant tissue explants (leaves, petioles), immature tissue	Gene integration into genome; multiple or partial copy insertions, typically requires regeneration and selection (species dependent)	Amenable to large sizes of DNA, rapid protocol pre-delivery, can deliver proteins	Low transformation efficiency, specialized equipment needed, large amount of DNA needed, tissue/cell damage	Day 1: sterilization of macrocarriers, holders Coating of carriers with DNA Particle bombardment²⁵
NP-mediated passive delivery	MSNs¹²	Mature tissue (roots), protoplasts	Fate of delivered DNA (genome integration versus transient expression) remains uninvestigated	High DNA-loading capacity	Unknown target species range (validated only in Arabidopsis)	Days 1 and 2: synthesis of MSN Day 3: functionalization of MSN Preparation of DNA-MSN complexes Plant incubation with DNA-MSN
	Peptide carriers^28,34	Mature tissue (leaves), immature tissue (embryos), targeted delivery to mitochondria^35,36	Fate of delivered DNA (genome integration versus transient expression) remains uninvestigated	Rapid protocol, no specialized equipment needed, no external force required	Unknown target species range (validated only in Nicotiana benthamiana and Arabidopsis thaliana)	Day 1: preparation of peptide-DNA constructs Infiltration into target tissue
	SWNTs^18,22	Mature tissue (leaves^18,22, roots and callus (data not shown)), protoplasts^18,22, for nuclear^18,22 or chloroplast²² expression	Transient expression of gene^18,22, no integration into genome¹⁸	No transgene integration, scalable, no external force required	Unknown target species range (validated in tobacco, arugula, wheat, cotton¹⁸, Arabidopsis thaliana²², sorghum (data not shown)	Day 1: COOH-SWNT suspension PEI-SWNT reaction Day 2: PEI-SWNT washing Preparation of PEI-SWNT + DNA Infiltration into/exposure to target tissue