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In vitro regeneration of sweet Gourd (cucurbita moschata Duch.)

M. Rezwan Kabir
Biotechnology Division, Bangladesh Agricultural Research Institute, Gazipur-1701, Bangladesh

M. A. Yousuf Akhond
Biotechnology Division, Bangladesh Agricultural Research Institute, Gazipur-1701, Bangladesh

M. Al-Amin
Biotechnology Division, Bangladesh Agricultural Research Institute, Gazipur-1701, Bangladesh

M. Shahidul Haque
Department of Biotechnology, Bangladesh Agricultural University, Mymensingh, Bangladesh

Abstract/ Executive Summary:

Cotyledon explants of sweetgourd (Cucurbita moschata Duch.) were cultured on MS supplemented with different concentrations of BAP alone and in combination with NAA, TDZ and 2,4-D. High shooting frequency (94.4%) was obtained from BARI Mistikumra-1 when cotyledon segments were cultured on MS with 4.44 μM BAP alone. The highest number (8.9) of shoot development and minimum days (14.7) required for shoot induction were also observed in 4.44 μM BAP treatment for BARI Mistikumra-1. In contrast, 83.3% shooting frequency were observed from BARI Mistikumra-2 producing 11.9 shoots in 8.4 days on 8.88 μM BAP. The regenerated shoots of both varieties were rooted on half strength of MS and MS supplemented with 0.53 μM NAA and 2.46 μM IBA. Rooting was observed in BARI Mistikumra-1 with 92.6% frequency in 12.3 days whereas it took 8.6 days with 93.3% rooting frequency in BARI Mistikumra-2 when shoots were cultured on MS supplemented with 0.53 μM NAA. Welldeveloped rooted plantlets were transferred to pots containing sterile soil and covered with polythene bags in the greenhouse for hardening. The acclimated plants were planted in the field after three weeks.

Keywords: Cotyledon, Cucurbita moschata, Regeneration, Sweetgourd

Location:

Start Date:

End Date:

Program Area: Crop-Soil-Water Management

Commodity: Sweet gourd

Objectives:

To establish an efficient regeneration protocol using two popular sweetgourd cultivars aiming at future genetic transformation.

Methodology:

Fresh, uniform and healthy seeds of sweetgourd cultivars BARI Mistikumra-1 and BARI Mistikumra-2 were collected and sterilized for 12 -15 minutes in 1% (v/v) sodium hypochlorite containing 3 - 4 drops Tween 20 in a laminar air flow cabinet. Afterwards, they were rinsed 4 times with sterile distilled water at 5 minutes interval and placed in test tubes containing half strength of MS with 30 g/l sucrose, vitamins and 8.5 g/l agar. The pH of the medium was adjusted to 5.8 and autoclaved at 121°C, 15 psi for 20 min. Initially the cultures were kept in dark for germination and after two days, transferred into a growth chamber maintained at 24 ± 2°C under a 16/8 hrs (light/dark) photoperiod. Cotyledons were carefully excised from in vitro grown seedlings and placed on sterile Petri dishes or hard papers. Each cotyledon was cut off close to the hypocotyl and then cut across in half with the distal parts discarded as the distal end of cotyledons was not found to produce any shoots in a previous study. The proximal parts were cut in half with each part having a part of the stalk attached. The apical bud of the seedlings was removed carefully from the explants. Explants of both the cultivars were placed on MS without growth regulators (control) and with different concentrations of BAP (2.22, 4.44, 6.66 and 8.88 μM), BAP and NAA in combinations (4.44 + 0.53, 8.88 + 0.53, 22.20 + 0.53 and 0.44 + 5.30 μM), TDZ (0.23, 0.45, 2.25 and 4.50 μM) and 2,4?D (2.26, 4.52, 6.78 and 9.04 μM), respectively. The explants were subcultured in MS having five different concentrations of BAP for further multiplication. Four explants were placed on each Petri dish (15 × 90 mm) containing about 30 ml medium. The Petri dishes were sealed with Parafilm and kept at 25 ± 1°C under a 16/8 hrs (day/night) photoperiod with a light intensity of 1500 lux and sub-cultured every 14 days on the same medium and kept under the same conditions. When shoots attained a height of 2.0 - 2.5 cm, they were cleaned, excised and transferred to half strength of MS, MS supplemented with 2.46 μM IBA and 0.53 μM NAA for root induction. Well developed rooted shoots were transferred to pots in sterile soil and enclosed with polythene bags to maintain high humidity. The plantlets were kept in the greenhouse and watered once or twice in a week while keeping in covered conditions. After 2 - 3 weeks, bags were removed and the plantlets were transferred to larger pots for further growth. The experiment was set up in a CRD with three independent replicates. The analysis of variance for different parameters was performed and the means were compared by R programme using STAR software at 5% level of significance.

Source of Information: Plant Tissue Cult. & Biotech. 26(1): 67?75, 2016 (June)

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

The regenerated shoots of both varieties were rooted on half strength of MS and MS supplemented with 0.53 μM NAA and 2.46 μM IBA. Rooting was observed in BARI Mistikumra-1 with 92.6% frequency in 12.3 days whereas it took 8.6 days with 93.3% rooting frequency in BARI Mistikumra-2 when shoots were cultured on MS supplemented with 0.53 μM NAA. Welldeveloped rooted plantlets were transferred to pots containing sterile soil and covered with polythene bags in the greenhouse for hardening. The acclimated plants were planted in the field after three weeks.

Knowledge Management: Journal