Bud Induction of Serpent Gourd (Trichosanthes anguina L.) In Vitro

Cucurbit Genetics Cooperative Report 23:80-82 (article 27) 2000

Zhang Lihua, Cheng Zhihui, Cui Hongwen and Xue Wanxin
Department of Horticulture, Northwest Science and Technology University of Agriculture and Forestry, Yangling, Shaanxi, 712100 China

Introduction. Serpent gourd belongs to Cucurbitaceae and originates in the tropic area of Asia. The cultivation of serpent gourd occurs mainly in India and other countries of southeastern Asia. In china, little serpent gourd has been grown though, cultivation of this plant has been increasing. Serpent gourd is nutritional and has high value as a vegetable. there are only a few seeds in each fruit and the propagation coefficient is very low. The cost and shortage of seeds has been the limitation to the enlargement of serpent gourd production. Therefore, development of a fast propagation system is a significant aid to increasing the cultivation of this vegetable.

Material and Methods. The healthy seeds of the serpent gourd cultivar endemic to the Xi’an area were scalded at 60 C for 10 min and soaked at 20 C for 48 hr. The seeds having broken seed coats germinated at 27-28 C and 2 to 3 days later, planted into blocks filled with sterilized compost. The blocks were placed in a room at 27+ 1 C, and 12 to 14 hr of 1500 to 2000 Lux of light.

Apex buds and axillary buds of seedlings were used as explants. The explants were cut into pieces 0.5 to 0.8 cm long and sterilized with 70% alcohol for 30 seconds, rinsed 3 time with sterilized water, then sterilized with 0.1% HgHCl2 for 3 min and rinsed 3 times again. The explants were planted on media following sterilization.

MS medium was used as the basal medium and supplemented with 2.5% sucrose and 0.6% agar.

Different concentrations of TDZ, KT, ZT, 6-BA, and various combinations of 6-BA with 2,3-D or IAA were supplemented to media for bud induction.

Results and Discussion. Effects of cytokinins on bud induction: Apex buds of 4-day-old seedlings were used as explants. Explants planted on the media supplemented with TDZ tended to produce callus. There was no bud initiation on the media supplemented with KT (0.1 or 2.0 mg/L) or TDZ (0.2 or 2.0 mg/L) or ZT at lower concentrations (0.05 or 0.2 mg/L). KT and ZT at lower concentrations only promoted explant elongation and root initiation, with the effect of KT greater than of ZT. Buds were initiated by ZT at higher concentrations (0.05 or 2.0 mg/L. However, the explants tended to elongate and the quality of initiated buds was poor. Buds were initiated at all concentrations of 6-BA tested (Table 1). The optimal 6-BA concentration was found to be 1 mg/L.

Effects of combinations of 6-BA with 2,4,-D or IAA:Apex buds and axillary buds of 3-week-old seedlings were used as explants. Buds were initiated in all treatments combining 6-BA and IAA. Similar bud initiation effects were found on the media supplemented with only 6-BA. However, greater IAA concentrations resulted in longer bud length (Table 2).

The explants tended to produce callus but no buds on media supplemented with 6-BA and 2,4-D at higher concentrations (0.1 or 0.2 mg/L) (Table 3). when lower concentrations of 2,3-D (0.05 or 0.02 mg/L) were combined with 6-BA, buds were induced. Differences in bud induction were also observed between explant sources. The apex and bud explants initiated buds only at the lowest concentration (0.02 mg/L) of 2,4-D in combination with either concentration of 6-BA. The axillary bud explants initiated buds at 2,4-D concentrations of 0.02 mg/L or 0.05 mg/L combined with either 6-BA concentration. Although buds were induced with combinations of 2,4-D and 6-BA, they were of poorer quality than those induced on the media supplemented with only 6-BA (Table 3).

Table 1. Effects of 6-BA on bud induction of Serpent gourd.

Concentration of 6-BA (mg/L)	Number of buds per explant	Bud length (cm)
0.5	4.3	2.1
1.0	5.3	1.1
2.0	too small to count	–

Table 2. Effects of combinations of 6-BA with IAA on bud induction of Serpent gourd.

Concentration of hormone (mg/L)		Explant	Number of bud per explant	Length of bud (cm)
6-BA	IAA
0.5	0.1	apex bud	2.5	0.87
0.5	0.2	apex bud	2.7	1.13
1.0	0.1	apex bud	3.7	0.51
1.0	0.2	axillary bud	3.4	0.70
1.0	0.1	axillary bud	3.5	0.38
1.0	0.2	axillary bud	3.7	0.41
1.0	0.5	axillary bud	3.8	0.47
1.0	0.8	axillary bud	4.0	0.52
1.0	1.0	axillary bud	3.7	0.64

Table 3. Effects of combinations of 6-BA with 2,4D on bud induction of serpent gourd.

Concentration of hormone (mg/L)		Explant	Number of bud	Length of bud (cm
6-BA	2,4-D
0.5	0.02	apex bud	2.0	0.3
		axillary bud	1.5	0.3
0.5	0.05	apex bud	*	*
		axillary bud	2.0	2.5
0.5	0.1	apex bud	*	*
		axillary bud	*	*
0.5	0.2	apex bud	*	*
		axillary bud	*	*
1.0	0.02	apex bud	5.5	3.4
		axillary bud	2.0	0.35
1.0	0.05	apex bud	*	*
		axillary bud	2.0	0.2
1.0	0.1	apex bud	*	*
		axillary bud	*	*
1.0 .	0.2 .	apex bud	*	*
		axillary bud	*	*

* Represents callus production of explants and no bud induced.

Literature Cited

1. Cao Huaxing, Gu Yongqiang, He Xiaohua. 1986. The effects of different cytokinine on the formation of adventitious buds in culture of watermelon. Journal of Shanghai Agricultural College. 4(4): 181-185.
2. Tang Dingtai, Zhang Jinglan, Xu Guifang et al, 1980. The effects of plant hormone on callus formation and regeneration of plant in Cucumis melo L.) Journal of botany 22(2): 132-135.
3. Tang Shaohu, Liao Yingfen, Xu Rongcan. 1994. Medium selection and tissue culture of seedless watermelon. Journal of Southwestern Agricultural University. 16(6):540-542.
4. Tao Yamin, Zhang Zhigang, Ye Jian, Pan Chongguang. 1994. Effect of several physical-chemical factors on somaclone propagation with seedless watermelon. Journal of Shanghai Agricultural College. 12(1):48-51.
5. Wang Xiaosu, LiBuxun, Wang Guangdong, 1997. In vitro plantlet regeneration from hypocotyl of Sechium deule Swartz. Journal of Northwestern Agricultural University. 25(1):83-87.
6. Kim., Y.H. and Janick, J. 1989. Somatic embryogenesis and organogenesis in cucumber. HortScience. 2:702.