Introduction

Rapid Mass propagation Techniques of Cassava Planting Materials for Nursery Establishment

Cassava ( Manihot esculenta Crantz.) is the most important rootcrop in the tropics and ranks fourth after rice, wheat and corn as calorie source for human consumption. In the Philipines it is utilized as food, feed, alcohol and the emerging biofuel industry. In support to the countrys cassava industry, PhilRootcrops undertakes a continuing crop improvement program to develop new and improved varieties. There are 46 high yielding varieties recommended for cultivation (2009). The demand for cassava in the country is not met. This is mainly due to the lack of planting materials of the high yielding varieties (HYVs) and the lack of capability to mass propagate the HYVs. These varieties do not reach the target clienteles because of slow and limited distribution of planting materials. Traditionally, cassava is propagated vegetatively using 15-30 cm cassava stakes with 7-8 viable nodes. Multiplication rate 1:10; one stem cutting will yield 10 planting materials after 10-12 months. Rapid propagation techniques are already available. It is essential to develop skills and capability for the mass propagation of cassava in order to sustain the supply of HYVs planting materials for industry. The brochure will show the rapid propagation techniques and production of clean planting materials for cassava. 1

A. Production of 2-node Cuttings

1. Select physiological mature, pest-and disease-free cassava stems. Cut from hardwood portion of the stems leaving at least 30 cm of the plant standing from the ground.

A mature cassava plant ready for production of 2 node cuttings

3. Pre-sprout the mini-stems in nursery beds or in perforated polyethylene bags filled with good quality soil. Prepare a nursery bed 1 meter wide of any convenient length under shade. Plant the two-node mini-stem horizontally in a small furrow in the bed at very close spacing but the mini-stems must not touch each other. Plant tip-shoot cuttings vertically and group them together in the nursery bed. If the polyethylene bag is used, plant the node cutting vertically position with two-thirds of the nodes buried in the soil. Place the planted polyethylene bags in partial shades. Follow the steps for nursery maintenance indicated below.

2. Cut the stems into mini-stems containing two viable nodes using a hack saw. Separate the hardwood or lower portion, the semi-mature or middle parts cuttings and the tip-shoot mini-stem cuttings. Dip the mini-stems in broad spectrum insecticides and fungicides to destroy the existing pests and diseases of cassava materials.

Mini stems planted in seedbed (A) and in polyethylene bag (B) Water the cuttings immediately after planting; and then either in the morning or late in the afternoon until 2 weeks after planting or when the plants have already established. Label the beds indicating the variety and planting date. Weed regularly to keep the nursery clean. Cover the cuttings that become exposed during watering Fertilized the plants with urea at the rate of 1.5 g/m2 Within one week after planting to ensure germination and vigorous growth.

Segregate mini stems; hardwood, middle part and tip shoot

4. Mini-stem cuttings (especially hardwood and semi-mature) sprout about 7-10 days after planting. Fibrous roots develop at the nodes and the oldest ends of the cuttings. Later, shoots emerge and develop leaves. Keep ministem cuttings in the nursery for 4-6 weeks before transplanting in the field.

6. After transplanting, maintain the field in order to produce strong, healthy plants. During the first 10 weeks, hoe to loose the soil and control weeds. Apply fertilizer where necessary and as appropriate. 7. At 8 months after transplanting, cut the stems and distribute to farmers. Do not uproot plants to harvest storage roots.

5. Transplant cuttings in a well prepared field at a spacing of 0.75 m-1 x 05.75 m. Firm the soul around each plant. Label the plots indicating the variety, date of planting and other important data.

Eight-month old cassava plants 8. Cut the stems at a height of 30 cm above the ground. Be sure that the stems are physiologically mature and free of pest and diseases. 9. Tie the stems together in bundles of desired number. Handle the stems with care when harvesting, loading, transporting and unloading to avoid bruising. Bruised buds may never develop into shoots.

Newly transplanted pre sprouted 2 node cuttings in the field 4

Bundled cassava stems ready for distribution 5

The practice of leaving stumps standing after cutting the stem is known as ratooning. Several shoots may sprout from each stump, but leave only three or four shoots. Apply fertilizer to the rationed fields.

10. Cassava cuttings cannot be stored for a long time because stems dehydrate. Storage can only be up to 8 weeks. When storing, place the stalks in an upright position (bud facing up) in a well ventilated shed or under a tree providing good shade.

The rationed field 2 months after cutting the stem

Other information

Cut another set of stems again 6 months later. Harvesting of stems could be up to three times.
Cassava yield using two-node cassava cuttings and traditional sets is comparable.

B. Tissue Culture
Stage 1 Donor Plant Established and Explants Isolation

Stage 2

In Vitro Stockhouse Establishment

Prepare modified Murashige and Skoog (1962) culture medium and sterilize using autoclave / pressure cooker for 15 minutes at 15 psi.

Newly established cassava plants in the screenhouse

Preparation of explants in the sterile chamber (A), Cultures maintain in incubation shelves (B)

Plant cassava stems cuttings in pots with sterilized soil and maintain under the screen house. Water the plants regularly and spray fungicides twice a week using the recommended rate. Apply insecticides if necessary. Harvest sprouts with 4-5 nodes from the tip.

Isolate single nodal explants inside the sterile chamber and inoculate immediately into a culture medium. Maintain the cultures in shelves illuminated with fluorescent tubes. The room temperature must be maintained at 25 + 2 C.

Newly harvested cassava sprouts (A), Disinfecting solutions (B) Remove the expanded leaves and petioles of the sprouts. Wash with tap water, soak in 2 % NaOCI for 5 minutes. Rinse the tissues with sterile distilled water three times inside the sterile chamber. 8

A- 1 week old B- 2 week old C-3 weeks old

In Vitro stock plants at different growth stages

Stage

Micropropagation

Plant the hardened cassava in the field at least 1 month after potting out. Follow the proper cultural management practices.

Micropropagated Cassava nodal explants

Hardened cassava plantlets ready for yield transplanting

After 1 month of incubation, 3-4 in vitro nodal stock plants are ready to sub-culture using single nodal explants. Complete plantlets are ready for plotting out after about a month from inoculation.

Stage

Potting-out and Field Maintenance

Transfer the cultures to ambient room condition for about a week before potting Transplant the plantlets in bottles to pots or plastic bag with sterile soil medium and place in the greenhouse for 3-4 weeks for hardening stage.

Cassava plantlets potted for hardening in the screenhouse

Comparison of the size of the traditional planting materials (A) and the first cycle tissue cultured derived planting materials planting materials (B)

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Yield in weight and number of storage roots in tissue culturedderived planting materials was higher in the genotypes studied.

Sources of Information and Pictures

Centro Internacional dala Agricultura Tropical (CIAT) Thailand and Cali, Colombia

Cassava Genotypres
Graphical presentation of the comparison between the tissue culture-derived planting materials and the traditional practice. Data reflected is for first cycle evaluation only.

Center for tropical Crops Research Institute (CTCRI) Trivandrum, India

Philippine Root crop Research and Training center (PhilRootcrops) Visayas State University, Visca, Baybay City, Leyte, Philippines

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