Sugarcane tissue culture protocol pdf

This is typically facilitated via the use of a liquid, semisolid, or solid growth medium, such as broth or agar, in vitro under sterile growing conditions. All media, supplement and reagents must be sterile to prevent microbial growth in the cell culture. Development of protocol is an iterative and research oriented process. There are certain basic steps for the regeneration of a complete plant from an explant cultured on the nutrient medium fig.

Production of healthy plant materials by shoot tip. Protocol optimization and histological analysis of in vitro. Tissue culture is the cloning of plants from mother plant. Brazil is the first largest world sugarcane producer country, with an estimated annual production of 690 millions of tons produced in approximately four million hectares fao, 2012.

Pdf an efficient protocol for large scale production of sugarcane. Tissues from mother plant are taken out and grown in nutrient media following the protocol. The experiments included four varieties, 9 callus induction media, 27 regeneration media, and 9 root induction media under twofactor. Some reagents and supplements will require filter sterilization. Plant tissue culture is a tool for obtaining rapid, mass multiplication of disease free, true to type planting material. Optimization of protocols for in vitro regeneration of sugarcane. It is the main sugar producing crop that contributes more than 75% to the total sugar pool at the global level. Optimization of protocols for in vitro regeneration of. Recent advances in sugarcane biotechnology, especially marker assisted. However, high costs involved in micro propagation are. Meristem tip or shoot tip culture using apical meristems has been used as a tool to produce virusfree plants hendre et al.

Improved tissue culture conditions for the emerging c 4 model. Somaclonal variation has potential to enhance genetic variation present within a species. Excellent multiplication response of in vitro formed shoots was obtained. Request pdf developing an efficient protocol through tissue culture technique for sugarcane micropropagation the present paper deals with the effect. A protocol for invitro direct plant regeneration from leaf tissues for. Tissue culture induced somaclonal variation limits the applications of tissue culture in sugarcane which owing to its high ploidy level is very unstable in vitro. Diseasefree production of sugarcane varieties saccharum.

Micropropagation of sugarcane through leaf sheath culture. Improved genetic transformation of sugarcane saccharum. An efficient protocol for micropropagation of sugarcane from shoot tip explants has been described. Evaluation of tissue culture raised crops of sugarcane as. 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. Present study was done to optimize an in vitro propagation protocol for sugarcane. Pdf protocol optimization for in vitro mass propagation. Plant tissue culture lab practices made easy for beginners. Plant differentiation from callus tissue of saccharum species. Medicinal plants are also being experimented by tissue culture method and. Production of healthy plant materials by shoot tip meristem. Sugarcane micropropagation using light emitting diodes and. Plant tissue culture is a practice used to propagate plants from.

Currently a production part of the sugarcane plantlets are produced by tissue. The following is a general guideline for culturing of cell lines. Development of protocols for invitro direct plant regeneration from. Virendra kumar jain, for always being there with me as celestial bodies. Jan 19, 2018 production of healthy plant materials by shoot tip meristem culture is a scientific and technical movie, illustrating the different stages of the meristem culture, that allows the regeneration. Mridula jain, who is an inspiration for me and have taught me to work hard with sincerity and devotion. An efficient protocol for micropropagation of sugarcane using. An exper im ent was conducted to devlop an efficient protocol for cropropagation of sugarcane using leaf sheath cultur method of three sugarcane varieties viz. This protocol has been used to successfully transform a commercial sugarcane cultivar, sp80. An efficient in vitro hardening technique of tissue. Panicum hallii vasey halls panicgrass is a compact, perennial c4 grass in the family poaceae, which has potential to enable bioenergy research for switchgrass panicum virgatum l. Assessment of genetic fidelity of sugarcane plantlets raised through. Plant tissue culture laboratory, northwest strategic technologies center cetene, cidade universitaria. Plant suspension cultures provide both faster growth than tissue culture and the ability for simple production, isolation, and purification of foreign proteins.

An efficient protocol for large scale production of sugarcane through micropropagation article pdf available in pakistan journal of botany 401. Pdf an efficient protocol for large scale production of. Mammalian cell tissue culture techniques protocol abcam. However, there was no data on the magnitude of the. In ethiopia, a lot of sugarcane tissue culture protocols were developed which help to minimize the current challenges of sugarcane production in the country. The experiments included four varieties, 9 callus induction media, 27 regeneration media, and 9 root induction media under twofactor factorial crd. Optimization of protocols for in vitro regeneration of sugarcane saccharum officinarum. Therefore, application of plant tissue culture techniques provides an. The aim of present study is to findout the meristem size of sugarcane varieties related to diseasefree with a high recovery percentage. Pdf protocol optimization for in vitro mass propagation of.

Therefore, the present investigation was undertaken to ensure that large numbers of clean explants should survive sterilization. This lecture also explains about the environment required for plant. Protocol optimization and histological analysis of in. The project entitled standardization of protocol for large scale production of sugarcane co86032 through micropropagation was thus undertaken to study and understand important growth parameters of invitro regenerated sugarcane plants in order to standardize tissue culture protocol for large scale production of sugarcane. Recife, pernambuco, brazil corresponidng author e mail. Technical laboratory activities on the production of sugarcane plantlets. In vitro plantlets of four sugarcane varieties, khonkan1, k9729, kps94 and k84200, were cultured on modifiedms media. Low cost options for tissue culture technology in developing. Commercial tissue culture industry is in existence globally since the last 25 years. Production of healthy plant materials by shoot tip meristem culture is a scientific and technical movie, illustrating the different stages of the meristem culture.

Shoot tip culture was considered a better method for micropropagation, since it produced plants phenotypically similar to the mother plant and gave a much more rapid multiplication rate when compared to the. Currently sugarcane production in ethiopia has given due attention, it is also allied with its important potential for an environmentfriendly biofuel production and generating huge. Tissue culture strategies in sugarcane saccharum officinarum. Shoot regeneration from inoculated shoot tip explants 4060% was obtained on ms medium containing bap, kinetin and ga 3 0. Suspension cultures allow for the generation of clonal variation within a single genotype more quickly than tissue culture.

Developing an efficient protocol through tissue culture. Tissue culture technique was used to obtain salt tolerant variants from embryogenic calli of sugarcane saccharum sp. Tissue culture is the growth of tissues or cells separate from the organism. Sugarcane tissue culture initiation studies gave a better knowledge about the sugarcane virus elimination under invitro conditions about the medium and various chemical constituents 9. The present study was initiated to optimize in vitro protocol for mass propagation of two commercial sugarcane clones co 449 and co 678 grown in ethiopia through shoot tip culture. Protocols for plant tissue culture tissue culture involves several stages of plant growth, callus formation, shooting, roots, and flowering. By plant tissue culture new plants may be raised in an artificial medium from very small parts of plants, such as, shoot tip, root tip, callus, seed, embryo, pollen grain, ovule or even a single cell, whether the cultured tissue develops into a plant or grows unorganized depends on the genetic potential of the tissue and the chemical and physical environment. This is due to regenerates has to adjust to many abnormalities in ex vitro environment like high level of irradiance, low humidity and water is. These are cultivated using excellent quality basic ingredients and advanced technology in adherence to set quality standards. Oct 18, 2016 tissue culture raised sugarcane crops showed 32% and 8% more cane yield in plant cane and first ratoon respectively. Sugarcane contributes 6070% of annual sugar production in the world. Sep 08, 2016 outcome the project in vitro regeneration of sugarcane saccharum officinarum l.

Steps on the working protocols of metahara sugarcane tissue culture laboratory. Hence, methods of more efficient regeneration protocol for propagation have. The invention discloses a tissue culture and rapid propagation method of sugarcane, and relates to the technical field of plant tissue culture. In vitro studies of sugarcane variety co91017 through. Dec 09, 2015 this video lecture explains about the process of plant tissue culture including the stepbystep procedure of micro propagation. Request pdf developing an efficient protocol through tissue culture technique for sugarcane micropropagation the present paper deals with the effect of light intensity, photoperiod and growth. Pdf in vitro direct plant regeneration using shoot tip. 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. Pdf sugarcane saccharum officinarum l tissue culture. Outcome the project in vitro regeneration of sugarcane saccharum officinarum l. Cell culture basics handbook thermo fisher scientific. In other words, it is an in vitro culture of plant cells or tissues on artificial nutrient media under aseptic conditions, in glass containers. Modifications of new cells to form tissues or organs with a specific function.

Cane and sugar yield of tissue culture plants similar to conventionally propagated plants. Doc steps on the working protocols of metahara sugarcane. Fundamental techniques in cell culture sigmaaldrich. Considering its importance in the agroprocessing industry and its economic contribution to the countrys, the ethiopian government has planned an ambitious plan. Tissue culture strategies in sugarcane saccharum officinarum l. In sugarcane after a long pursuance and hectic experimentation we successfully developed protocols for tissue culture propagation of almost all the popular sugarcane varieties in pakistan. Explant tissue taken from its original site and transferred to an artificial medium. The method comprises the following steps of selecting sugarcane axillary buds. Plant tissue culture of sugarcane linkedin slideshare. Micropropagation has been extensively used for the rapid multiplication of many plants species. Plant tissue culture, cell culture or micropropagation is the technique of.

Ex vitro rooting of sugarcane saccharum officinarum l. Our range of products comprises banana tissue culture plant, sugarcane tissue culture plant, ornamental and other plants such as aloe vera, ficus, heliconia, xanadu, dieffenbachia, spathiphyllum etc. 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. In vitro direct plant regeneration using shoot tip explants in sugarcane saccharum officinarum l. Present investigation indicates the in vitro regeneration of sugarcane explant possible with hormone. Tissue culture offers an opportunity to mass produce diseasefree planting material and is now used to supplement commercial sugarcane propagation in many countries including brazil, the united states, india corresponding author. Improved tissue culture conditions for the emerging c 4. Although many countries are sugarcane producers only six of them account for 65% of the world entire sugarcane production, brazil being the largest one. Optimization of sterilization protocol for two sugarcane varieties viz. An efficient protocol for commercial plant production. It starts with sterilizing a piece of plant tissue. The present investigation is an attempt towards finding an effective sterilization protocol for two, field grown sugarcane varieties, cos 88230 and cos 767 from meerut region and to establish the callus culture protocol. An efficient protocol for micropropagation of sugarcane. Early assessment of genetic fidelity in sugarcane saccharum.

Suspension cultures can be synchronized to obtain a homologous population of. Considering the above advantages, micropropagation has an important role to play in the seed production chain in suga rcane. Center for horticulture biotechnology, directorate of research, university of horticulture sciences, bagalkot 587 104, karnataka state corresponding author email. Plant tissue culture offers the best methodology through micro propagation of sugarcane for quality and phytosanitary planting material at a faster rate in a shorter period. Technology uses meristem to clone the mother plant. Request pdf tissue culture strategies in sugarcane saccharum officinarum. Shoot tips were surface sterilized with 5% active chlorinated berekina for 25. Protocol optimization for in vitro sugarcane establishment.

Experiments on shoot multiplication and rooting were laid out in a completely randomized design with factorial treatment arrangements. Cn104106467a tissue culture and rapid propagation method. All cell culture must be undertaken in microbiological safety cabinet using aseptic technique to ensure sterility. Shoot tip culture was considered a better method for micropropagation, since it produced plants phenotypically similar to the mother plant and gave a much more rapid multiplication rate when compared to the other procedure. Cp48103 that were cultured on a selective medium containing different. Standardizing sterilization protocol and establishment of.

Culture conditions culture conditions vary widely for each cell type, but the artificial environment in which the cells are cultured invariably consists of a suitable vessel containing a substrate or medium that supplies the essential nutrients amino acids, carbohydrates, vitamins, minerals, growth factors, hormones, and gases o 2, co. The developments in sugarcane tissue culture have already boosted sugarcane improvement to a great extent. The handbook is intended as a guide rather than an indepth text book of cell culture and you are encouraged to consult relevant specialised literature to. About 18, 520 plants, produced from a single shoot. Isd16, isd36 and isd37 in the germplasm laboratory of breeding division. Also the success of the sugarcane tissue culture is also dependent on. Wahome abstract the purpose of this study was o develop an alternative, reliable and economical sugarcane propagation system using tissue culture techniques. With this view, the ethiopian sugar industry uses this technology since 2012. Later intensive work about sugarcane development by using the above technique was amended byliu 10 byinculcating. Plant tissue culture lab practices made easy for beginners 2016 4 acknowledgement i am thankful to god almighty and my dad mr. Technologies for cost reduction in sugarcane micropropagation. A tissue culture technique in sugarcane indian institute of sugarcane research. Sugarcane is globally the main source of raw material for the production of sugar. Sugarcane tissue culture plant ramrich biotech private.

A protocol for a particular variety is a detailed procedure to multiply it. Tissue culture can increase the propagation potential by 2035 times 15. General technique of plant cell, tissue and organ culture is almost the same with a little variation for different plant materials. Tissue culture techniques have been widely used in saccharum spp. Review on in vitro propagation of sugarcane to advance the. Microprop g tion of sugarcane through leaf sheath culture. However, its wider use is restricted often by the high percentage of plant loss or damaged when transferred to ex vitro condition.

Sugarcane saccharum officinarum l is commercial cash and sweet industrial crop. The meristem culture is the only method to remove the scmv sugarcane mosaic virus as the meristematic tissue remains free from virus disease. Tissue culture is an exciting new way to rapidly produce and supply diseasefree seed cane of existing commercial varieties. Green shoots are generally observable within three weeks, and roots develop within six weeks. Tissue culture technology, if properly incorporated into the system of sugarcane production, has better rate of propagation, more cane and sugar yield than the conventional seed source and hence makes the sugar industry more profitable.