@ Mohamed,

1. I think they are trying for 'transient' assay with the Maize leaves. The message you mentioned (up to 50% transformation efficiency) was used for 'stable' transformation and published. The material used was the immature embryos.

2. For your question, I think they just want to do some cellular localization and interactions, not mass production of a specific recombinant protein. The possible reasons that they chose 'Maize' as experimental material can be that the promoter used in the construct is a monocot promoter, or simply they just want to see the results in Maize. Otherwise, they can also conduct particle bombardment on Onion (monocot) epidermis cells for transient assay.

I do most Agroinfiltration on Tobacco leaves, not on the Maize leaves. Below message is just my opinion:

I think that (1) it will be difficult to push the Agrobacterium culture into the Maize leaves which have a monocot leaf structure with a syringe (see attached picture; from website). (2) Besides, in general, Agrobacterium works more efficient towards dicot plants, not monocots (although nowadays immature embryos can be routinely used for transformation with a good transformation frequency). Particle bombardment can be an alternative method for gene transient assay.

The critical issue here is the low transformation efficiency of Agrobacteria for monocots. Tobacco and a number of other Dicots are very easy, but you'd probably need to look through the literature which strain, plasmid etc.. are suitable. To get the bacteria into the leaf is in my opinion not the problem, either by injection or by vacuum infiltration. Be careful with autofluorescence and use a positive control (e.g. GFP targeted to the nucleus). You can also think of delivering two genes on the same T-DNA, using  one marker to highlight transformed cells and another one for your gene of interest to do the localization and interaction studies. Alternatively, you may want to look into particle bombardment, which is probably the most widely used method for monocots.

If none has conducted Agroinfilteration in maize, the question should be why?. What are the advantages of conducting it on maize even if you are going only to optimize the conditions or to be the first one to report that. Maize is monocot, yes under certain conditions it can be transformed with Agrobacterium and the transformation frequency can be increase by supplementing Acetosyringone. But, what are the advantages of Maize leaves for mass productions of a certain component or protein, why maize leaves? 

If you would like to transiently express some proteins in maize to investigate their sub cellular localization, yes it can be done. Maize is transformable up to 10% or more with Agrobacterium, the tissue and the genotypes will play a significant role in transformation frequencies. Make sure you use one of the most common Agrobacterium tumefaciense strains like LBA4404, EHA101 or AGLO... they are widely used with monocots; sorghum and maize. Also, make sure you supplement Acetosyringone in the infection process to aid and increase the transformation frequency of a reluctant crop for Agro-mediated transformation. I hope you can find a previous report, if no I wish you a good luck optimizing the conditions

Sorry the transformation some times can be as a great as 50%. So please make sure you work on a highly transformable genotype like A188 

Stage of leaf development can als be considered a factor here along with specific genotype, most virulent strain and phenolic inducing compounds Acetosyringone, MES etc

@ Mohamed,

1. I think they are trying for 'transient' assay with the Maize leaves. The message you mentioned (up to 50% transformation efficiency) was used for 'stable' transformation and published. The material used was the immature embryos.

2. For your question, I think they just want to do some cellular localization and interactions, not mass production of a specific recombinant protein. The possible reasons that they chose 'Maize' as experimental material can be that the promoter used in the construct is a monocot promoter, or simply they just want to see the results in Maize. Otherwise, they can also conduct particle bombardment on Onion (monocot) epidermis cells for transient assay.

@Yuan-Yeu Yau @Mohamed

Yuan, you are right. We just want to confirm a protein-protein interaction in maize after we spray with different hormones. We choose maize because we are working with maize proteins and I think is nicer to make the confirmation in the same plant specie.'

Thanks,

Jorge

Hey all! I am also looking for transient expression system in maize. I need to test sub-cellular localization of candidate proteins. Bring on suggestions for the best way to go. 

Thanks

Hardeep

@ Hardeep,

Can you try particle bombardment if a machine is available in your university, because you just want to test sub-cellular localization.