Abstract
Floral initiation and development in the angiosperms is the essential step on which the yield of the plant depends. Sometimes external climate or any abiotic stress hinders the floral initiation and ultimately affect the plant yield. Hence, in vitro floral induction and development can help to overcome the external climatic factor. Furthermore, the protocol for in vitro floral induction in the parasitic angiosperm like Cuscuta reflexa has not been reported yet. We have standardized the protocol for floral induction in the parasitic plant Cuscuta reflexa. In this study it is established that MMS (modified Murashige Skoog) media supplemented with 2 mg L-1 NAA (naphthalene acetic acid, plant growth regulator) and 2 mg L-1 2,4-D (2,4-dichlorophenoxy acetic acid, plant growth regulator) supported floral induction after shooting in vitro. Furthermore, we found that MMS media supplemented with 2 mg L-1 2,4-D rapidly induced floral buds directly from the nodal explants without any shoot elongation. This protocol will help the researcher to induce flower in vitro in the other angiosperm plants along with Cuscuta reflexa.
Keywords: Cuscuta reflexa, in vitro, Floral induction, NAA, 2,4-D
Background
Cuscuta reflexa is a parasitic angiosperm parasitizing on a huge number of angiosperms (Kuijt, 1969). The ability to cause severe damage and loss of yield in the host plant has made this species important for scientific study (Nun and Mayer, 1999). Most of the Cuscuta species are non chloroplastic except few which have functional chloroplasts (Hibberd et al., 1998). Development of seedlings from larger embryos of C. reflexa in white’s medium has been reported (Maheshweri and Baldev, 1961). Floral induction of C. reflexa in short day period as well as dark conditions has been seen in vitro (Baldev, 1962). In vitro floral induction of Cuscuta japonica in short day conditions has also been established by Furuhashi et al. (1991).Specifically, floral induction and effect of growth regulators on floral induction of C. reflexa has not yet been studied. A complete tissue culture system for Cuscuta trifolli in liquid MS culture has been reported so far (Bakos et al., 1995). The floral induction in C. reflexa on modified white’s medium, subjecting the plant to different light and dark conditions, has been studied years back (Baldev, 1962). Here we have shown a complete in vitro culture system for floral induction in C. reflexa. Concentration of 2,4-D played a significant role in floral induction. Supplementation of NAA along with 2,4-D induces shoot followed by flower, but supplementation of only 2,4-D induces flower directly from nodal explants without showing any stem elongation. This result is very attractive and is showing the importance of 2,4-D in floral induction of this plant.
Materials and Reagents
Equipment
Procedure
Data analysis
The experiment has been repeated thrice independently. All the analysis has been done manually or visually.
Notes
Recipes
Note: All the solutions can be freshly prepared or stored solutions (at 4 °C, stable for 30 days).
Acknowledgments
Authors are thankful to Department of Botany, Utkal University, Govt. of Odisha for the financial support. Authors acknowledge previous work published by Das et al. (2011), from which this protocol is adapted.
References
If you have any questions/comments about this protocol, you are highly recommended to post here. We will invite the authors of this protocol as well as some of its users to address your questions/comments. To make it easier for them to help you, you are encouraged to post your data including images for the troubleshooting.