1.

Transgenic crops with improved quality traits

(A) Flavr Savr Tomato

• ‘Flavr Savr Tomato’ was the first genetically engineered crop granted license for human
consumption and commercialization.
• It was produced by Calgene Inc., Davis, USA in the year 1992.
• Calgene won FDA approval for Flavr Savr on May 18, 1994. Three days later (May 21,
1994), Flavr Savr tomato (the genetically engineered tomato) was introduced under the
brand name MacGregor at a pair of grocery stores—one in Davis, CA, USA and the
other in Northbrook, IL, USA - each of which sold more than 3,000 pounds of the GM
tomatoes.
➢ In tomato, enzyme polygalacturonase (PG) degrade pectin, which is the major
component of fruit cell wall.
➢ PG is responsible for the softening of fruits and deterioration in food quality.
➢ Calgene developed transgenic tomato (known as Falvr Savr Tomato), which
contain antisense construct of the gene (reverse-orientation copy of the gene)
encoding PG.
➢ The transgenic plants (Flavr Savr tomatoes) show a drastically reduced expression
of PG and significantly slower fruit softening.
Antisense RNA technology:
o In any gene, the DNA strand oriented as 3’→ 5’ in relation to its promoter is transcribed;
this strand is called the antisense strand.
o The mRNA base sequence, therefore, is complementary to that of the antisense strand.
o The remaining DNA strand of the gene, called sense strand, is naturally complementary
to the antisense strand of the gene.
o Therefore, the base sequence of the sense strand of a gene is the same as that of the
mRNA produced by it (except for T in place of U). Hence, the hnRNA / mRNA produced
by a gene in normal orientation is also known as sense RNA.
o An antisense gene is produced by inverting, i.e. reversing the orientation of, the protein-
encoding region of a gene in relation to its promoter. As a result, the natural sense strand
of the gene becomes oriented in the 3’→5’ direction with reference to its promoter, and is
 transcribed. The RNA produced by this gene has the same sequence as the antisense
strand of the normal gene, and is, therefore, known as antisense RNA.
o When an antisense gene is present in the same nucleus as the normal endogenous gene,
transcription of the two genes yields antisense and sense RNA transcripts, respectively.
Since the sense and antisense RNAs are complementary to each other, they would pair to
produce double-stranded RNA molecules. This event makes the mRNA unavailable for
translation.

A schematic representation of antisense RNA technology (Source: B.D.Singh)

Use of antisense RNA technology for production of Flavr Savr Tomato

Calgene used antisense RNA technology against PG enzyme encoding gene. On the basis
of PG gene sequence, an antisense PG gene was constructed and transgenic tomato plants were
created. These transgenic plants produced both sense and antisense mRNA for the PG gene
resulting in RNA-RNA pairing. This will not allow the PG gene to express, and the
nonproduction of PG enzyme prevented the degradation of pectin of the tomato fruit cell wall
and in turn preventing the softening of fruits.

(Source: H.S.Chwala)
Or
[Production of Flavr Savr (transgenic tomatoes) using antisense RNA approach; Source: Internet]

➢ Thus, these Flavr Savr tomatoes have about 2 weeks longer shelf-life than normal
tomatoes.
➢ Most importantly the Flavr Savr tomatoes have improved flavor and total soluble
solid (TSS), in addition to the enhanced shelf life. These desirable features are
due to the fact that the fruits of Flavr Savr can be allowed to ripen on the plants.
(Due to extra shelf life the Flavr Savr tomato can transported to market after vine-
ripening). Thus, the fruits accumulate more sugars and organic acid to improve
their taste and TSS content. In contrast, the conventional tomatoes picked green &
hard and are treated with ethylene during storage to ripen; thus without full array
of vine ripened tomato flavours.
➢ The research and marketing efforts that produced the Flavr Savr tomatoes resulted
in a scientific success, a temporary sales success and then commercial demise.
A number of reasons are being given for the failure of Flavr Savr tomatoes. (1)
The demand of the Flavr Savr Tomato was high, but the product was never
profitable because of high production and distribution cost, (2) The transgene was
put into some very poor variety with relatively poor yield, (3) Public opinion and
perception about use of genetically modified food.