Plant Tissue Culture Techniques for rapid growth of Bamboo
Different propagation techniques are available for bamboo such as seed propagation, clump division, rhizome and culm cuttings. These classical techniques suffer from serious drawbacks for large or mass scale propagation.
Given below is a brief about the large scale micro propagation on bamboo using techniques like micro propagation, somatic embryogenesis, in vitro flowering, macro proliferation along with quality control parameters like field performance, clonal fidelity.
Micro propagation:
Extensive research on micro propagation of bamboo species has been carried out using
A. Juvenile (Zygotic embryo, seed or seedling)
B. Mature clump derived (nodal buds) tissues with more than 40 species of bamboo.
Micro propagation from Seedlings
Advantage:
Use of a new generation as well as easier technique for in vitro multiplication.
Disadvantage:
A. Insufficient knowledge of genetic background
B. Restricted availability of seeds
C. Loss of germination capacity etc.
Micro propagation from mature tissue explants
In order to supplement conventional methods of propagation, an efficient in vitro propagation method by using explants from selected mature plants would offer a desirable alternative for large-scale propagation of bamboo.
Advantage:
Micro propagation through axillary bud proliferation by nodal explants is the best method to maintain the clonal fidelity. Being a clonal method, it highly reduces or eliminates the variation inherent in seed raised population
Disadvantage:
But, endogenous contamination and instability of multiplication rates affect the flow of in vitro mass propagation.
Limitation:
This tissue culture protocols from mature tissues of bamboo were very limited to only few species
Technique 1 : Induction of Roots
Roots were induced on shoots within 20 to 45 days of explanting period either on auxin or on a combination of both auxin and cytokinin enriched medium.
Points to Note:
Apart from optimum concentration of growth regulators, selection of appropriate size of shoot propagule selected from a healthy multiple shoots having 1 to 2 cm in length
Longer shoots (> 2.0 cm) with folded leaf lamina showed a lower rooting percentage.
Placing of single shoot in rooting media, failed to induce root formation. Three shoots (1 to 2 cm long) was the best for root induction of D. asper.
Technique 2 : SOMATIC EMBRYOGENESIS
Tissues which were not committed to an embryogenic pathway can be induced to have embryonic determination by exposure to an auxin.
Benefits:
Somatic embryogenesis can provide a convenient and dependable source for obtaining plants for bamboo.
Points to note:
To get somatic embryonic callus, explant selection is the most crucial factor. Callus have been raised from various explants, viz. seeds, seedlings, isolated embryos, roots, shoot apices, leaf explants, internodes and anthers.
Technique 3 : IN VITRO FLOWERING
Flowering in bamboo is a biological enigma that happens at long infrequent intervals, in every 2 to 3 times in a century
There are few hypothesis
a. Drought creates competition among the shoot clumps for survival, for which flowering may be formed for generation of offspring and this triggers mast flowering
b. Bamboo produce large quantity of seeds and storage of food reserve takes long time
c. Since bamboo plants die after flowering may be genes that are involved in programmed flowering followed by cell death.
In vitro flowering can open up the possibility of controlled flowering and develop viable spikelet’s and produce fertile seeds
Technique 4: MACROPROLIFERATION
By cutting the rhizome into pieces, each with roots and shoots, each seedling can be multiplied three to seven times depending on species.
Generally, this method is only suitable for species producing seeds.
After successful acclimatization, macro proliferation can be very suitably adapted for well-established bamboo plantlets after three to five months of transfer of the micro plants. By splitting the rooted tillers, it was possible to increase the production up to three times.
This process can be continued for a number of years. By recycling of the macro-proliferation procedure, continuous plantlet production resulted.
Benefits:
A. Proliferated plantlets were small in size, hence easy to handle and transport.
B. A small initial stock could produce large numbers of plants
FIELD PERFORMANCE OF MICROPROPAGATED BAMBOO
The tissue-culture raised bamboo plantlets are being assessed in terms of:
A. Height of the plant
B. Diameter and number of culms produced
Results are:
A. Bamboo culm formation was observed within one year in micro propagated plants compared with two years in seedling-derived plants.
B. In plantlets derived from tissue culture, the height of the culm, the number of culms per plant, the number of nodes of the main culm, and girth of the second internodes were nearly double those of seedling-derived plants
Clonal Fidelity
The way to determine the somatic clones derived from tissue culture can be done through molecular markers came with polymerase chain reaction (PCR) technology with Random identified polymorphic DNA (RAPD)
Why clonal fidelity check is required?
It is necessary to define the plant quality standard to establish an accurate and economic way.
A suitable quality control strategy is necessary to ensure the performance of the bamboo plants and the genetic pool of the planting stocks.
The lack of reports on ascertaining the genetic fidelity of tissue culture raised bamboo could lead to an undesirable variant lasting for several years.
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