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Morphological Change Due to Effects of Acute Gamma Ray on Wishbone Flower (Torenia fourmieri) In Vitro

Morphological Change Due to Effects of Acute Gamma Ray on Wishbone Flower (Torenia fourmieri) In Vitro

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*Corresponding author (
Anchalee Jala
). Tel/Fax: +66-2-5644440-59 Ext. 2450. E-mail:anchaleejala@yahoo.com. 2011. International Transaction Journal of Engineering,Management, & Applied Sciences & Technologies. Volume 2 No.4. ISSN 2228-9860.eISSN 1906-9642. Online Available athttp://TuEngr.com/V02/375-383.pdf 
 
375
International Transaction Journal of Engineering,Management, & Applied Sciences & Technologies
 
http://www.TuEngr.com,http://go.to/Research 
Morphological Change Due to Effects of Acute Gamma Ray onWishbone Flower (
Torenia fourmieri
)
 In Vitro
 
Anchalee Jala
a*
 
a
 Department of Biotechnology, Faculty of Science and Technology, Thammasat University,THAILAND
 
A R T I C L E I N F O
 
A B S T RA C T
 Article history
:Received 17 June 2011Received in revised form01 August 2011Accepted 03 August 2011Available online03 August 2011
Keywords
:Wishbone Flower,Gamma Rays,Acute Irradiation,Morphology,Tissue Culture
 
Young shoot tips were used as explants and cultured onMS medium supplemented with varying concentrations of (0.1,0.5, 1.0 mg/l) BA and (0.1, 0.5, 1.0 mg/l) NAA. Calli and newshoots were grown on MS medium supplemented withcombination of 0.5 mg/l BA and 0.5 mg/l NAA. New shootsaveraging 0.5–0.8 cm were irradiated with varying doses of gamma rays (5, 10, 15, 20 grays). Gamma irradiation hadvarious effects on growth of 
Torenia fournieri
. Higher dosage of gamma irradiation reduced plant height, number of roots, number of leaves, leaf length, leaf width, petiole length and number of guard cells at abaxial and adaxial epidermis surface. Plantmorphology and flower development was also modified.
2011 International Transaction Journal of Engineering, Management, &Applied Sciences & Technologies. Some Rights Reserved.
 
1.
 
Introduction
 
Wishbone flower or 
Torenia fournieri
is a member of the Scrophulariaceae family. It isgenerally a perennial plant which is normally grown as an annual shrub. With an approximateheight of 12 inches, it is preferably planted along with many other similar species to ensure awidespread flowering bed. The plant looks like a nice little green shrub. The mature plant isdensely branched and decorated with shiny green leaves and delicate cup-shaped flowers. It isgrown as a pot plant or used for decorating and landscaping. Demand for the plant has
 
2011 International Transaction Journal of Engineering, Management, & Applied Sciences & Technologies.
 
 
376
Anchalee Jala
continued to increase. Induced mutation has been reported to be an efficient technique toachieve the desirable characters in flowers and ornamental plants (Maluszynski, 1995).Gamma rays generally influence plant growth and development by inducing genetic, biochemical, physiological, morphological and anatomical change in cells and tissues (Gunckeland Sparrow, 1961). Various effects of gamma rays on ornamental plants areobserved in the different generations after mutation induction. M1 generation isheterogeneous with different mutations for different plants. It also exhibits non-heritabledirect effects on mutagens such as sterility. Chimeric heterozygous at mutations are change of genetic material that may be transferred from M1 to the following generations (Gaul. H.,1977).The objective of this study was to use an
 In Vitro
mutation technique to improvewishbone flower in order to select suitable colors for growing and to study morphologicalchange after transplanting to soil.
2.
 
Materials
 
and
 
Methods
 
Young shoot tips of wishbone flower were used as explants materials. These explantswere sterilised with 5.25% calcium hypochlorite and cultured on MS medium (Murashige andSkoog, 1962) containing 30 g/l sucrose, 2.5 gm/l gelrite and supplemented with varyingconcentrations (0, 0.1, 0.5 mg/l) BA (Benzyl adenine) and (0, 0.1, 0.5, 1.0 mg/l) NAA(Naphthaline acetic acid). After calli were formed and multiplying shoots developed, theculture was irradiated with varying doses of gamma ray (0, 5, 10, 15, 20 grays). Followingirradiation, M
1
V
1
shoots were immediately cut into small pieces, each piece had 2 nodes withaverage length of 0.5-0.8 cm and subcultured into fresh medium with the same formula(MS+0.5 mg/l BA and 0.5mg/l NAA) at 4 weeks interval from M
1
V
1
to M
1
V
4
. All thesewere maintained at 25º ± 1ºC under 16 hr cool white, fluorescent light (1600 luxs)(Dooley,1991). The plants were transferred to soil to observe their growth.Data collection was undertaken of plant height, number of roots, root length, number of leaves, leaf length, leaf width, petiole length, leaves arrangement on node. Guard cells fromabaxial and adaxial epidermis surface were examined by light microscope. Each slide wasrandomly sampled to determine guard cell frequency by using images viewed under 
 
*Corresponding author (
Anchalee Jala
). Tel/Fax: +66-2-5644440-59 Ext. 2450. E-mail:anchaleejala@yahoo.com. 2011. International Transaction Journal of Engineering,Management, & Applied Sciences & Technologies. Volume 2 No.4. ISSN 2228-9860.eISSN 1906-9642. Online Available athttp://TuEngr.com/V02/375-383.pdf 
 
377
magnification of 10x40.
3.
 
Results
 
After young shoot tip explants had been cultured on MS medium with six differentconcentrations of BA and NAA for 8 weeks, the effective results were obtained as shown inTable 1. After one week some explants swelled, turned green, and Calli were formed and proliferated new shoots within 8 weeks. Samples which cultured in 0.5 mg/l BA and 0.5 mg/l NAA (Figure 1a) were the best. This was followed by a combination of 0.1mg/l BA and 1.0mg/l NAA. MS medium supplemented with 0.5 mg/l BA and 0.5 mg/l NAA was used for multiplication of new shoots (Figure 1b).
Table 1:
Callus induction from shoot tip explants of 
Torenia fournieri
cultured on MSmedium with combinations of NAA and BA for 8 weeks.
MS medium Number of callus
a
 Visual Observationof callusShoot formation NAA(mg/l)BA(mg/l)Shootheight(cm) No. newshoot(shoot) No.nodes/plant(node)0 0 1 swollen - - -0.1 0.1 1.6 Small and creamy 1.2 0.9 1.20.5 0.1 2.1 Medium and creamy 1.17 2.2 1.40.5 0.5 4.1 Big and light green 1.12 3.4 2.41.0 0.1 2.6 Medium and light yellow 1.10 2.1 2.21.0 0.5 1.7 Medium and light brown 1.16 1.2 1.6
a- Callus growth was graded by an index of 1 – 5: 1 - indicating no callus formation3- indicating medium sized, and 5 - indicating the biggest sized of callus formation
Figure 1:
Effects of BA and NAA in MS medium on calli induced and shoot regenerated onwishbone flower explants:(A): calli induction, (B): shoot regeneration, (C) : young shoots before irradiated gamma rays New young shoots (0.5-0.8cm) (Figure 1C) (M
1
V
1
) irradiated with gamma rays weresubcultured fourfold (M
1
V
4
) in the same medium every 4 weeks. The elongated shoots (about
A BC

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