Micropropagation of an elite F1 watermelon (Citrullus lanatus) hybrid from the shoot tip of field grown plants

2022

HBR

The current study was carried out to elucidate a reproducible protocol to develop plants directly from leaf and internodes to facilitate the genetic transformation in Coccinia cordifolia (L.) Cogn., a medicinal plant of the cucurbitaceae family. In vitro grown leaf and internodes were used, which were collected for regenerated shoots from field-grown nodal segments that were sterilized by 0.1% HgCl2 treatment for 6 minutes. The nodal segment cultured on BAP, Kn and BAP, NAA combination where 1.5 mg/L BAP solitary supplement and augmented supplement with 0.1 mg/L NAA was most effective as 80% shoots regenerated with 4.0±0.37 and 2.7±0.45 shoots per culture, respectively. Collected leaf and internode responded 90% at 1.5 mg/L BAP + 0.1 mg/L NAA fortified full-strength MS medium. The highest number of shoots also regenerated in the same combination which were 8.1 ± 0.30 and 10.2±0.40, respectively and found internode as the best explant for direct organogenesis. For root induction, half strength of MS medium supplemented with 0.1 mg/L IBA was found most effective. The highest number of roots regenerated per shoot (6.8±0.10) and root length (2.8±0.20). The successful acclimatization of the in vitro (80%) grown plantlets proved the validity of the developed protocol of using biotechnological techniques for improving the plant.

Section: Discussionmentioning

confidence: 61%

Direct in vitro organogenesis from leaf and internode of Coccinia cordifolia (L.) Cogn.

Sarkar¹,

Rahman

2022

HBR

“…Sekhawat et al ( 2014) also reported the potentiality of half-salt strength in root formation and observed the best result in 2.0 mg/l IBA. Sundari et al (2011) also reported IBA as an effective auxin for root formation, similarly IBA supremacy for root induction has also been reported in many others species of Cucurbitaceae (Hoque et al 2007, Khalekuzzaman et al 2012, Arciniega-Carreon et al 2017. Well rooted plantlets were transferred into a thumb pot containing sand : garden soil : organic fertilizer (1:2:1) for hardening.…”

Section: Resultsmentioning

confidence: 76%

Indirect Organogenesis from in vitro Derived Leaf and Internodes of Coccinia cordifolia (L.) Cogn. - An Important Medicinal Climber

Sarkar¹,

Plant Tissue Cult. & Biotech.

2022

An efficient in vitro regeneration protocol was developed via callus from the explants of leaf and internode of Coccinia cordifolia (L.) Cogn., an important medicinal climber. Among the different combinations of auxins and cytokines used, BAP with NAA were found highly effective for induction of callus from in vitro derived leaf disc and internode explants. Maximum callus formation rates were recorded on MS with 1.0 mg/l BAP and 0.1 mg/l NAA for both leaf disc and internode explants. The best organogenesis via callus was obtained on MS with 2.0 mg/l BAP and 0.5 mg/l NAA. The highest numbers of shoots regenerated per culture were 6.5 ± 0.10 and 5.6 ± 0.55 for leaf disc and internode explants respectively and leaf discs was found to be better for indirect organogenesis. For root induction, half strength of MS medium supplemented with 0.1 mg/l IBA was found to be the most effective. The mean number of roots regenerated per shoot was 6.8 ± 0.10 with an average root length of 2.8 ± 0.20 cm. In vitro derived plantlets were acclimatized successfully. This protocol for in vitro regeneration can be utilized for the improvement of this climber through biotechnological methods. Plant Tissue Cult. & Biotech. 32(2): 127-136, 2022 (December)

“…In vitro clonal propagation has been one of the most using technology for plant preservation and rapid propagation (Anisuzzaman et al, 2008). So, the tissue culture technology is widely exercised for large-scale reproduction and protection of genetic resources of medicinal plants and have been successfully applied to cucurbitaceae plants (Pal et al, 2007;Khalekuzzaman et al, 2012;Alam et al, 2019).…”

Section: Discussionmentioning

confidence: 99%

Propagation of Coccinia cordifolia (L.) Cogn. from shoot tip and nodal segment through micropropagation techniques

Sarkar¹,

Rahman

2021

HBR

The present work was undertaken to develop a reproducible protocol for the micropropagation of an important medicinal plant of the Cucurbitaceae family, Coccinia cordifolia (L.) Cogn., by using shoot tips and nodal segments to overcome the impediment in seed settings and seed germination of conventional reproduction. To develop an efficient protocol, 0.1% HgCl2 treatment for 6 minutes was found effective for surface sterilization of field-grown explants to eliminate microbes and fungus and to get healthy tissues. Throughout the study, different concentrations of auxin, cytokine, and gibberellin were used either alone or in combination as supplemented in MS medium to find suitable conditions and suitable explant (nodal segment). BAP was the best cytokine source as 2.0 mg/l BAP produced 4.0±0.20 and 4.8±0.09 shoots per culture and gained 5.8±0.11 cm and 6.7±0.32 cm length with regeneration rate of 90 and 100 percent in shoot tips and node, respectively. The highest percentage (90%) of regeneration for axillary shoot proliferation was obtained in node explants at MS medium containing 2.0 mg/l BAP + 0.1 mg/l NAA. The highest number of shoots regenerated per culture was 3.0±0.41, with a length of 6.0±0.16 cm. GA3 was also found effective in producing longer shoots with the combination of BAP, but the average shoot number was reduced drastically. Although full-strength MS medium was found to be ideal for shoot regeneration and used in shoots proliferated experiments, half and full-strength of MS medium with auxins supplements of 0.5, 1.0, and 2.0 mg/l either of IBA, NAA, and IAA) were used for roots growth and half strength nutrients supplemented with 0.5 mg/l IBA was found most compelling. The highest number of roots regenerated per shoot was 3.1±0.30, and the average root length was 1.8±0.30 cm. Rooted plantlets were finally transplanted into small plastic pots containing sun sterilized sand, soil and humus (1: 2: 1) to adapt the plantlet in ex vitro environment, and acclimatized plantlets showed 95% survival rate in outdoor condition which proved the effectiveness of using biotechnology to improve plant’s growth rate and mass production.