Freshly harvested roots of cassava varieties were peeled, washed and then crushed separately. Cassava plantlets covered with polystyrene cups and placed in a water bath containing high phosphorus fertiliser. Plant regeneration via protoplast electrofusion in cassava. Cell cluster formation by leaf protoplasts from axenic cultures of haploid petunia hybrida l. In vitro studies on cassava plant micropropagation of cassava manihot esculenta crantz book pdf available. Plant tissue culture, cell culture or micropropagation is the technique of producing selected plants of known desirable agriculture qualities, in large numbers of plants from small pieces of plant in relatively short period times. Micropropagation of manihot esculenta crantz cassava. Hardening procedure for banana the procedure for the hardening of tissue cultured banana plantlets is. In vitro micropropagation of cassava through low cost tissue culture. Where permitted, plants in tissue culture can be safely exchanged if certified diseasefree.
Temporary immersion system and hardening of massive numbers of cassava vitroplants bernardo ospina1, roberto segovia2 and armando bedoya3 abstract tissue culture is a technique widely used for propagation of plant material. The disposable petri dishes and baby food jar were inoculated with three cassava nodal cuttings each, while the test tube was inoculated with one cassava nodal cutting. Tissue culture laboratory, punjab agricultural university, ludhiana, punjab india. When we wish to reuse the stored pgr we have to use the tissue culture method to propagate it. A definition of meristematic tissue in plant biology. Cassava production and limitation of propagation through tissue culture. Cassava is a perennial shrub with a height of 1 to 5 m, palmate leaves with 5 to 9 leaf lobes, and it is grown. This eliminates disease pressure otherwise caused by transfer of cuttings. Cauliflower florets in particular give excellent results since they can be grown into a complete plant in the basic tissue culture media, without the need for additional growth or root hormones. Plant cell and tissue culture a tool i n biotec hnology ba sics.
An improved protocol for the culture of cassava leaf protoplasts. Recently, it has been demonstrated that acetone cyanohydrin is. Effect of light intensity and orientation on invitro. Cassava starch as alternative lowcost gelling agent for in. A two factorial experiment was conducted with the first factor as type of gelling agent 80 gl cassava flour. Going forward, we hope to provide technical guidance for cassava tissue culture, and also provide some useful inspiration and reference for further genetic improvement of cassava. Use of tissue culture in cassava for rural households in. This is to minimize the spread of the pests and systemic diseases of this important food crop. The rapidly growing tissue culture tc industry within east africa, aiming to provide planting material to banana farmers, has stimulated discussion about the need for virus indexing to. The genus manihot consists of at least 98 species rogers and appan 1973.
In vitro techniques for cassava conservation should be used whenever technical expertise, facilities and operating budget are available. In vitro propagation of malaysian cassava manihot esculenta. They are generally more economical and less risky in a longterm perspective, as compared to field collections. It was pointed 77 out that cassava flour at 8% wv provides optimum gelling. In this research, a low cost protocol for cassava tissue culture was developed and used to regenerate two farmer preferred cassava varieties, kme 1 and. We further analyzed the differences existing between each tissue derived. Modifications of new cells to form tissues or organs with a specific function. Meristem micropropagation of cassava manihot esculenta frontiers. Another important conclusion from this work is the confirmation of a viable protocol for the regeneration of plants from cassava protoplasts. Effect of in vitro culture on epigenetic differentiation between tissues. This study was conducted in the plant tissue culture laboratory of the agricultural research corporation arc during 2005 to 2008 to develop in vitro propagation techniques for cassava genotypes. Effect of various factors on minimal growth in tissue. However, production is constrained by lack of diseasefree planting materials.
Genetic stability of cassava plants regenerated through. The potential of a tissue culture medium gelled with cassava flour to support shoot proliferation of stem nodal sections of uapaca kirkiana and faidherbia albida was studied. Pdf cassava production and limitation of propagation. A system for vegetative propagation by stakes stems or propagules has therefore been developed to maintain traits of interest. Commercial tissue culture industry is in existence globally since the last 25 years. Costeffective nutrient sources for tissue culture of cassava. Plants important to developing countries that have been grown in tissue culture are oil palm, plantain, pine, banana, date, eggplant, jojoba, pineapple, rubber tree, cassava, yam, sweet potato, and tomato. This makes tissue culture an important technology in setting up cassava seed systems. In malaysia, cassava is an important industrial crop for starch processing and food industries. Tissue culture techniques for rapid propagation of.
Cassava manihot esculenta crantz belongs to the family euphorbiaceae. Tissue culture refers to the production of cassava plantlets resulting from the growth of cells. The problem of genotype stability tissue culture storage methods conservation of cassava germplasm by means of meristem culture section h. Tissue culture is seen as an important technology for developing countries for the production of diseasefree, high quality planting material and the rapid production of many uniform plants. References anthony p, davey m r, power j b, lowe k c.
Callus was not formed at the rootshoot transition zone. Cassava genetic transformation and its application in. Low cost tissue culture of selected cassava manihot esculenta. Cassava is a highly heterozygous crop and many cultivars do not flower or do not produce enough, viable seeds. Pdf an efficient and promising protocol for in vitro propagation of cassava plant manihot. Introduction cassava manihot esculenta crantz is one of the most important crops in the subsaharan africa. There is therefore, need to put in place interventions that will reduce the cost of production hence making tissue culture products affordable. This article describes the adaptation of conventional cassava propagation to a lowinput scheme for rural tissue culture multiplication, developed and run by small, resourcepoor farmers referred in this article as an informalfarmers seed production system. The effects of different nutrient media formulations gamborg et ai. Pdf in vitro micropropagation of cassava through low. Commercial seed sources there are no commercial seed sources for cassava in the u. Abstract defined media that promote the initiation and undifferentiated growth of callus derived from stem explants of four cultivars of cassava, manihot esculenta.
Osmotic concentration, due to presence of mannitol, was only effective if. A method is presented for the rapid in vitro propagation of cassava manihot esculenta. Cassava plants produced through our proprietary tissue culture and dipropagation technologies are clean from cassava mosaic disease cmd and cassava brown streak disease cbsd. For this method, explants tissue from the plant are obtained from an invitro derived plant and introduced in to an artificial environment, where they get to proliferate. This can be circumvented through tissue culture but the technology is costly limiting its adoption. There is therefore, need to put in place interventions that will reduce the cost of production hence making. Potential of cassava flour as a gelling agent in media for. Meristem micropropagation of cassava manihot esculenta. Use of tissue culture in cassava for rural households in colombia. In the cyanogenic crop cassava manihot esculenta, crantz, the final step in cyanide production is the conversion of acetone cyanohydrin, the deglycosylation product of linamarin, to cyanide plus acetone. Low cost options for tissue culture technology in developing. In this research, a low cost protocol for cassava tissue culture was developed and used to regenerate two farmer. In vitro propagation systems that are based on the use of microcuttings or meristem cultures are considered analogous to clonal cuttings and so widely viewed to be largely free from such somaclonal effects. Notably, the most recent introductions of germplasm to both rwanda and burundi, from the kenya plant health inspectorate service kephis plant quarantine station at muguga, nairobi, and supported by earrnet, have been in tissue culture form.
Tissue culture has been used to develop more than 1,000 plant species, many of which are used commercially. Unfortunately, tissue culture is labor intensive, time consuming, and can be costly. However, somaclonal variation is a common phenomenon in tissue culture which makes it mandatory to monitor the genetic stability of plants. Application of tissue culture technology is constrained by high costs making seedlings unaffordable. Tissue culture technology of cassava manihot esculenta crantz is a viable alternative to currently adopted techniques for mass propagation, germplasm conservation and genetic improvement. Cryopreservation is used for longterm storage of pgr. In this study, the malaysian cassava variety which was putih variety was cultured onto low cost tissue culture media by using nodal explants. Complete cassava plants and callus tissues have been obtained from. Pdf costeffective nutrient sources for tissue culture of.
A method is presented for the rapid in vitro propagation of cassava manihot esculenta crantz. Methods of cultivating cassava meristem tissue are described. Research open access proteome characterization of cassava. Micropropagation, which is a form of tissue culture, increases the amount of planting material to facilitate distribution and large scale planting. The potential of using biotechnology to improve cassava. The objective of this study was to evaluate the possibility of using locally available fertilizers as alternative nutrient sources for cassava. Therefore, the objective of this study was to evaluate the. Tissue culture cassava plantlets hardened and ready for planting plantlets generated by the tissue culture method will provide a large quantity of clean i. Single cassava plantlet, removed from tissue culture container. Green shoots are generally observable within three weeks, and roots develop within six weeks.
In vitro micropropagation of cassava through low cost tissue. Pdf in vitro micropropagation of cassava through low cost. Tms 970162, 920323, and tme 419 cultures were maintained at 28 c 2 illumination and 16h. Tissue culture techniques are requisite in the quest to improve cassava productivity in the subsaharan africa and globally. Plant cell and tissue culture a tool in biotechnology basics. Productive potential of cassava plants manihot esculenta. Biotechnology unit plant tissue culture lab of national root crops research. Tissue culture systems were developed for cultivars of cassava grown in both southern africa and australia. Tissue culture of cassava on chemically defined media parke. Plant tissue culture terminology differentiatedcells that maintain, in culture, all or much of the specialized structure and function typical of the cell type in vivo.
Genetic transformation of cassava manihot esculenta crantz. Shoot tip culture may be described as the culture of terminal 0. The major features of the culture medium to be considered are its nutrients, growth regulators and physical mature. Cultiyars responded differently to low temperature. Tissue culture has been effectively applied in elimination of viruses and other systemic diseases from cassava vegetative materials allowing exchange and conservation of rejuvenated propagation materials with higher yields jorge et al.
Murashige and skoog, 1962 called ms and schenk and hildebrandt. In vitro culture of cassava manihot esculenta crantz. Through the cassava genome hub, an online platform that produces and. Hence, an in vitro propagation technique is needed to produce these highly demand industrial crop.
In vitro micropropagated samples were obtained from the tissue culture facility of the mikocheni agriculture research institute mari dar es salaam, tanzania and were propagated as described by konan et al. However, in india, plant tissue culture industry started about 15 years back and is revolutionizing the commercial agriculture sector by enabling mass propagation of quality planting material. Using meristem derived in vivo plantlets and leaves from plants grown under environmentally controlled conditions, a method was established to isolate consistently high yields of viable protoplasts and a new medium promoting protoplast division was formulated. This process occurs spontaneously at ph greater than 5.
This method has been used successfully to propagate species such us cassava, sweet potato, plantain and sugarcane. Tissue culture of cassava manihot esculenta crantz. Starch preparation the cassava varieties used tms 970162, 920323 and tme 419 were obtained from nrcri, umudike. Seed culture is the type of tissue culture that is primarily used for plants such as orchids. Tissue culture techniques for rapid propagation of selected. Similarly, acedo and labana 2008 noted rapid propagation of released philippine cassava varieties through tissue culture, suggesting their genotypic specificity to culture medium.
In vitro micropropagation of cassava through low cost. Costeffective nutrient sources for tissue culture of. In this research, a low cost protocol for cassava tissue culture was developed and used to regenerate two farmerpreferred cassava varieties, kme 1 and. As for all crop species, transgenic systems in cassava are reliant on the development of tissue culture systems capable of generating totipotent cells and tissues 9. Explant tissue taken from its original site and transferred to an artificial medium. Hussey g 1978 the application of tissue culture for the vegetative. Cassava genetic transformation and its application in breeding f. Shoot tips and nodal segments excised from in vitro manihot esculenta cultivar. Nodal explants were induced to grow as multipleshoot cultures on a medium containing 1. Tissue culture, cassava, nutrient sources, micropropagation, low cost medium, acclimatization. In vitro propagation of cassava manihot esculenta crantz. Media routinely used in cassava tissue culture modi. The term is adapted from the greek word merizein, meaning to divide, a reference to the function of the cells in the meristematic tissue.
Application of biotechnology in plant disease management. Aug, 2015 there is great interest in the phenotypic, genetic and epigenetic changes associated with plant in vitro culture known as somaclonal variation. Biotechnology unit plant tissue culture lab of national root crops research institute nrcri umudike, abia state nigeria. Adoption of droughttolerant crops such as cassava can help alleviate food insecurity in subsaharan africa.
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