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In Vitro Slow-growth Conservation for Two Genotypes of Solanum Tuberosum L.

M Nasiruddin
Department of Botany, University of Rajshahi, Rajshahi-6205, Bangladesh

AKM Rafiul Islam
Department of Botany, University of Rajshahi, Rajshahi-6205, Bangladesh

Abstract/ Executive Summary:

To find out the most suitable culture medium formulation to induce slow-growth and reduce the frequency of sub-culturing of the in vitro conserved microplants of the two potato genotypes at 24 ± 1ºC was conducted. Growth was controlled by using different concentrations of sucrose, mannitol, sorbitol (30 g/l) alone or in combination with either mannitol (15, 20 and 25 g/l) or sorbitol (15, 20 and 25 g/l) in murashige and Skoog medium. The results showed that single treatment (Sucrose or mannitol or sorbitol alone) was not feasible for long-term conservation. Combined treatment was responding better and maximum microplant survived (80.82 - 83.15%) after 12 months of storage on (T-8) medium supplemented with 10 g/l sucrose and 20 g/l sorbitol. In this formulation microplants were in very good condition, without phenotypic abnormalities and had enough nodes for sub-culturing up to 12 months. Microplant survival and condition were closely associated with each other but not with root growth.

Keywords: Potato, Microplant, Slow-growth, Osmoticum, In vitro conservation, Solanum tuberosum

Location: Rajshahi, Bangladesh

Start Date:

End Date:

Program Area: Crop-Soil-Water Management

Commodity: Tomato

Objectives:

To conserve potato germplasm in vitro at 24 ± 1°C by employing different concentrations and combinations of metabolically inactive sugar- alcohols (osmoticums) to produce osmotic stress for reducing sub-culture time of the microplants.

Methodology:

Single nodal segments from in vitro grown shoots of two potato genotypes viz., Asterix and Diamant were used to conduct different experiments. These genotypes were selected for their contrasting response to in vitro minimal growth. Further, they have wider genetic base. After sprouting from potato disease free microplants were cultured and maintained on MS medium supplemented with 30 g/1 sucrose under standard culture conditions (16 hrs photoperiod, 40 µmol/m²/s light intensity and 24 ±1ºC). Single nodal cuttings (SNCs) from primary culture were sub-culture on MS medium supplemented with different concentrations of sucrose, mannitol, sorbitol (30 g/l) alone or in combination with either mannitol (15, 20 and 25 g/l) or sorbitol (15, 20 and 25 g/l). Every sub-culture vessels containing 20 ml of medium solidified with 7 g/l Nobel agar (Merck, India). Culture tubes were closed with polypropylene caps and sealed with parafilm M (Laboratory Film) (Chicago, II. 60631, USA), and incubated at 16 hrs photoperiod (From cool white fluorescent lamps, approx. 20 µmol/m²/s light intensity) at 24 ± 1ºC in tissue culture room. The experiment was conducted in factorial (9 medium × 2 cultivars) randomized complete block design with 6 replicate culture tubes which were used in this investigation. Microplant survival, microplant condition (on a visual 0 - 5 preference scale: 0 = dead, 1 = very poor, 2 = poor, 3 = moderate, 4 = good to 5 = very good) for suitability of sub-culturing, root growth (on a visual 0-5 preference scale: 0 = nil, 1 = very poor, 2 = poor, 3 = moderate, 4 = good to 5 = very good), shoot length (cm) and number of nodes per microplants data were recorded after 3, 6 and 12 months of incubation. Observations were also made on the presence or absence of aerial roots, microtubers or phenotypic abnormality. The data on percent microplant survival were transformed into arec sine and those of microplant condition and root development into square roots (√x + 0.5). Both non-transformed and transformed data used to similar results, so only non transformed data were analyzed. Analysis of variance and correlation coefficient were carried out according to the standard procedures (Gomez and Gomez 1984). Experimental values are given as mean, the mean were compared using Duncan’s multiple-range test (DMRT) as outlined by IBM SPSS software version 20 (SPSS Inc. USA).

Source of Information: Bangladesh J. Bot. 47(3): 369-380, 2018 (September)

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

The results of correlation coefficients showed that microplant survival percentage was highly and positively correlated with microplants condition score (r = 0.94) and number of shoots per microplant (r = 0.81). Microplant condition score also showed significant and positive correlation with number of shoot per microplants (r = 0.68) after 12 months of in vitro conservation . The method of slow-growth conservation in vitro at normal propagation temperatures has been used to conserve germplasm of potato (Solanum tuberosum L.). Therefore, the method followed in this investigation might be a very promising method and desirable not only to save on the cost of energy and maintenance etc., but also for better genetic stability of the germplasm. In vitro slow-growth storage techniques are being routinely used for medium-term conservation of numerous species, both from temperate and tropical origin, including crop plants, e.g. potato, musa, yam, cassava Engelmann (1999) and rare and endangered species. However, this method also has some problems yet to be resolved. Further, it has been grown awareness in the last decade and offer many advantages as a complement to field maintenance.

Knowledge Management: Journal