Transgenic Plants Suitable For Transport and Long Shelf Life, Bruise Resistant, Antisense RNA, Polygalacturonase

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Transgenic plants suitable for transport and long shelf life

A number of examples are available, where transgenic plants suitable for food processing have been developed.
(i) Bruise resistant tomatoes were developed which express antisense RNA against polygalacturonase (PG) that attacks pectin in the cell walls of ripening fruit and thus softens the skin.
(ii) Tomatoes and cantaloupe melon fruits exhibiting delayed ripening were developed, either by using antisense RNA against enzymes involved in ethylene production (e.g. ACC synthase or ACC oxidase), or using gene for ACC deaminase, which degrades 1 amino-cyclopropane-l carboxylic acid (ACC), an immediate precursor to ethylene.

This will increase the shelf life of tomato. The tomato and cantaloupe melons expressing antisense ACC synthase or ACC oxidase gene will exhibit delayed ripening, of fruits that are still attached to the plant. However, in tomatoes expressing ACC deamainase gene, fruits displayed delayed ripening when detached from the plant and allowed to ripen on the shelf.

When fruits are allowed to stay on the plant longer, they accumulate higher amounts of soluble sugars, thus achieving better flavour. Tomatoes obtained thus (using ACC synthase) were named 'Flavr Savr'.

Production of transgenic 'Flavr Savr' tomato using an antisense RNA gene

Production of Flavr Savr		Mechanism Involved
(a) a gene of tomato codes for polygalacturonase (PG) that causes the fruit to soften		(synthesis of mRNA for PG)
(b) an antisense gene for PG was isolated and sequenced		(synthesis of antisense mRNA for PG)
(c) an antisense PG gene (the new Flavr Savr gene) was constructed, cloned in bacteria and then transferred to tomato		since opposite RNAs pair, PG gen is not expressed

(d) transgenic Flavr Savr tomato with PG gene and antisense gen produced		Flavr Savr tomato

(iii) Tomatoes with elevated sucrose and reduced starch could also be produced using sucrose phosphate synthase gene. (iv)Tomatoes with high level of total solids and soluble solids were produced using isopentenyl transferase (ipt) gene from Agrobacterium tumefaciens. This is a key enzyme in the biosynthesis of plant growth regulator, cytokinin.

Expression of the gene was modulated using a promoter permitting ovary specific expression. This led to increase in cytokinin level in tomato ovaries giving higher level of solids. (v) Starch content in potatoes could be increased by 20-40% by using a bacterial ADP glucose pyrophosphorylase gene (ADP GPPase).

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