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Bioscience Methods 2012, Vol.3, No.4, 27
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33
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27
Research Report Open Access
Factors Affecting Regeneration of Tomato Cotyledons
Wei Zhang , Leiping Hou , Hui Zhao , Meilan Li
College of Horticulture, Shanxi Agricultural University, Taigu, 030801, P.R. China
Corresponding author email: limeilan_2@hotmail.com;
Author
Bioscience Methods 2012, Vol.3, No.4 doi: 10.5376/bm.2012.03.0004
Received: 02 Feb., 2012
Accepted: 25 May, 2012
Published: 07 Jun., 2012
This article was first published in Molecular Plant Breeding (2010, 8(5): 913-919) in Chinese, and here was authorized to translate and publish the paper in English
under the terms of Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work
is properly cited.
Preferred citation for this article as:
Zhang et al., 2012, Factors Affecting Regeneration of Tomato Cotyledons, Bioscience Methods, Vol.3, No.4 27
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33 (doi: 10.5376/bm.2012.03.0004)
Abstract
In this research, we employed the tomato cultivar named Zhongshu Si Hao as research material to study the influences on
the regeneration of adventitious buds under the different conditions with hormone combinations, various explants, different parts of
cotyledons and cotyledon inoculating methods; we also screened the best method for seed surface disinfection and the most
appropriate medium for the rooting of adventitious buds. The results showed that the best procedures for surface disinfection of
tomato seeds would be as follows: firstly, immersing the seeds in 70% ethanol for 30 second, then rinsing them in 20% NaClO with
tween
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20 for 20 min. The most ideal explant for inducing buds is the middle part of the cotyledon, followed by the stem and the leaf.
the best medium formula for cotyledon regeneration is MS+ZT 2 mg/L+IAA 0.1 mg/L, and placing the cotyledon upside down
is better to induce the adventitious buds than that placing upside. The best medium for rooting of adventitious buds is
1/2MS+IAA 0.3 mg/L. This research would lay a foundation establish transformation system of tomato in our future research.
Keywords
Tomato; Cotyledon; Hormone; Bud induction
Introduction
Tomato (
lycopersicum esculentum
Mill), an annual or
perennial plant, belongs to
Lycopersicon
genus in the
Solanaceae family; the tomato has become a worldwide
fruit and vegetable due to its beautiful appearance,
fragrant flavor, nutrition and high economic value,
especially in recent years with the discovery of
lycopene, anti-oxidation and anti-cancer function,
edible benefit and economic value of the tomato are
received more attentions (Liang and Wang, 2003,
Xinjiang Petroleum College of Education, 7(2):
28-30, 34).
In production, the tomato often suffered from the
pathogen infections of blight, fusarium wilt, gray
mold, late blight and early blight infection; some
varieties have little resistance resulting in yield
decrease. In recent years, the great success is
achieved in genetically engineered resistant gene
transferred to plants, these technologies have also
become an important means in tomato breeding
program (Gao, 2006).
At present, although it has been reported that the
exogenous gene could be introduced into the tomato
genome by pollen tube transformation (Sanford et al.,
1985; Huang et al., 1997; Tian et al., 2000), the most
popular and mature technology applying in the genetic
transformation of tomato should be
Agrobacterium
-
mediated method. A variety of exogenous genes have
been successfully tranferred into tomato by using
Agrobacterium
-mediated method (Pozueta-Romero et
al., 2001; Van Roekel at al., 1993; Frary and Earle,
1996).
However, the efficiency of
Agrobacterium-
mediated
transformation of tomato is known to be pretty low. It
the process of tomato genetic transformation, the
average efficiency of the cotyledon transformation
was only 7.6% based on several transformation tests
with triparental mating
Agrobacterium
(Ye, 1994); it
was reported that the regeneration frequency of
cotyledon for transforming ACC synthase gene and its
antisense gene were 9.8% and 12.7%, respectively
(Tang et al., 1994, Chinese Science Bulletin, 39(24):
2280-2283). No doubt, the most successful transformation
system is built on the better tissue culture system in
the field of plant genetic engineering.
There are lots of studies on tissue culture of tomato;
much more regeneration system have been established
(Ou et al., 2002; He et al., 2003), but it is obvious that