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Standardization of Micropropagation Protocol for Enhancing Large-scale Production of BARI Released Strawberry Varieties (2019--2020)

M. M. Khatun
Biotechnology Division, BARI, Joydebpur, Gazipur

D. Khanam
Biotechnology Division, BARI, Joydebpur, Gazipur

M.A.Y. Akhond
Biotechnology Division, BARI, Joydebpur, Gazipur

Abstract/ Executive Summary:

Shoot tips of BARI Strawberry-2 and BARI Strawberry-3 were cultured on MS medium supplemented with different concentrations and combinations of BAP, Kn and GA₃. Four different treatments T₁, T₂, T₃, & T₄, were tested for shoot multiplication and elongation. In case of BARI Strawberry-2, for shoot elongation 0.5 mg/l BAP + 0.5 mg/l Kn +1.5 mg/l GA₃ combination was found most suitable. Whereas in BARI Strawberry- 3, 1.0 mg/l BAP + 0.5 mg/l Kn +1.5 mg/l GA₃treatment was found most suitable for shoot formation.In rooting, ½MS medium without hormone produced maximum root than full MS medium in both varieties.

Keywords: Strawberry, Laminar airflow, Micropropagation

Location:

Start Date: 00-00-2019

End Date: 00-00-2020

Program Area: Variety and Species

Commodity: Strawberry

Objectives:

The present experiment is important to develop a suitable protocol for mass propagation of BARI strawberry varieties

Methodology:

BARI Strawberry-2 and BARI Strawberry-3 were collected from the fruits field of HRC, BARI and used as plant materials. Small shoot tip came out from runner were carefully removed by scalpel from Strawberry plants showing no virus symptoms. The explants were prepared by removing the outer layers of tissue from shoot tips by scalpel. The light greenish 2-3 cm shoot tips were surface sterilized with Ridomil Gold 68WG for 5 min. to remove the fungal contamination. Then shoot tips were sterilized with Jet and Trix under running tap water for 5-6 min followed by 3-4 times washing with distilled water. Thereafter, shoot tips were sterilized with Clorox solution (60 ml Clorox: 40 ml distilled water) with 2 drops of Tween-20 for 3 min. followed by 3-4 times washing with sterile distilled water inside the laminar airflow cabinet. The dead tissue of sterilized explants was cut off to leave a 1-2 cm portion containing an intact apex. The prepared explants were placed in MS media (Murashige and Skoog, 1962) with four concentrations (T₁, T₂, T₃, and T₄) and combinations of BAP, Kn and GA₃. All cultures were incubated at 25 ± 1⁰C with a 16 h photoperiod (2000 lux) provided by cool white fluorescent tubes. The pH of the medium was adjusted to 5.8. Data collected were analyzed using R version 3.5.0 statistical program.

Source of Information: Annual Research Report 2019-2020, Biotechnology Division, BARI, Joydebpur, Gazipur

Article Link (Online Journal):

Funding Source:

1.

Budget:

Total Budget (Tk.) :

Achievement/Findings:

Shoot tip explants of BARI Strawberry-2 and BARI Strawberry-3 were cultured on MS medium supplemented with four different concentrations and combinations of BAP, Kn and GA₃. Shoot initiation started within 2-5 days after inoculation. In BARI Strawberry-2, early shoot initiation was noticed from T₃ treatment (2.50 days) while in BARI Strawberry-3, minimum days (2.58 days) required for shoot initiation in T₄ treatment. In BARI Strawberry-2, significant variation was observed in shoot length and root number. The highest shoot length was obtained from T₁ treatment (6.27 cm) in MS medium and the lowest (4.09 cm) was obtained from the ½MS T₃ treatment. For shoot elongation, MS medium is found better than ½MS. In leaf number, the maximum leaf (8.42) was found in MST₄ treatment and the lower (5.42) was obtained from MST₃ treatment. In BARI Strawberry-3, a significant difference was observed in all parameters. The shoot length was recorded fromMST₂ treatment (5.38 cm) and the lowest (3.62 cm) was obtained from the ½MST₃ treatment. For shoot elongation, MS medium is found better than ½MS. The maximum leaf (8.08) was found in MST₃ treatment and the lower (6.25) was obtained from ½MS T₄treatment. In vitro grown shoots were cultured in both MS and ½ MS medium without hormone for rooting. Results showed that a hundred percent of shoots produced roots in all treatments. In BARI Strawberry-2, a significant difference was noticed among the different treatments. The shoots derived from T₂treatment cultured in ½MS medium produced maximum roots (8.25) followed by T₁ treatment in ½ MS medium (6.75). On the other hand, a lower number of roots (3.08) was observed from shoots of T₃treatment cultured in MS medium. The highest root length (3.08 cm) was noticed from shoots derived from T₁ treatment cultured in MS medium followed by ½ MS T₂ (2.98 cm) medium. Although, there is no significant difference between MST₁ and ½ MST₂ in root length. The lowest root length (1.73) was observed from shoots of T₃treatment cultured in ½ MS medium. In BARI Strawberry-3, the shoots derived from T₁treatment and cultured on ½ MS medium produced maximum roots (6.83) followed by T₂ treatment in ½ MS medium (6.58). Although, there is no significant difference between ½ MS T₁ and ½MS T₂ treatment in root number. On the other hand, a lower number of roots (3.92) was observed from shoots of T₄treatment cultured in MS medium. The highest root length (3.25 cm) was noticed from shoots derived from T₃ treatment cultured in MS medium (Table 3). The lowest root length (0.46) was observed from shoots of T₄treatment cultured in ½ MS medium. Results indicated that in strawberry rooting no growth hormone is needed. In both the varieties, results showed that for root formation, ½MS is better than full MS. Means bearing the same letters do not differ significantly at the 5% level of probability by LSD. Well-rooted plantlets in test tubes were kept at room temperature for 2 to 3 days for hardening. After that, the plantlets were removed from the culture tubes and all the adhering media were washed carefully so that, the root damage was the least. The plantlets were then planted into small plastic pots containing sand, soil and decomposed cow dung at the ratio of 1:1:1 and covered with polythene bags to maintain high humidity for one or two weeks. After four weeks they were transferred to big plastic pots. The plantlets were successfully established in potting soil and ready to be transplanted in the field.

Knowledge Management: Report/Proceedings