Abstract
Pigeon pea (Cajanus cajan (L.) Millsp.) is a drought tolerant pulse legume, mainly grown for grain in the semiarid tropics, particularly in Africa. Pigeon pea production in countries like Kenya is faced with a number of challenges, particularly lack of high quality seeds. The objective of this study was to develop an in vitro regeneration system for pigeon pea varieties grown in Kenya, that is amenable to genetic transformation. In vitro regeneration of pigeon pea varieties, KAT 60/8 and ICEAP 00557, commonly grown in Kenya was achieved using leaf explants from in vitro grown seedlings, through callus initiation, followed by shoot and root induction. For callus initiation, MS media supplemented with 0.5-4 mg 1-1 2, 4-D and TDZ separately were tested, and IBA at 0.1, 0.5 and 1 mg 1-1 was tested for rooting of shoots. Embryogenic calli was obtained on MS containing 2, 4- D; whereas TDZ induced non-embryogenic callus alone or with shoots directly on explants. Indirect shoot regeneration frequency of 6.7 % was achieved using 1 mg 1-1 2, 4-D-induced embryogenic callus obtained using KAT 60/8 explants. Whereas direct shoot regeneration frequencies of 20 and 16.7% were achieved using ICEAP 00557 and KAT 60/8 explants, using 0.5 mg l-1 and 2 mg 1-1 TDZ, respectively. Optimum rooting was achieved using 0.5 mg 1-1 IBA; and up to 92% rooted shoots were successfully established in soil after acclimatisation. Genotype and hormone concentrations had a significant (P<0.05) influence on callus, shoot and root induction. The protocol developed can be optimised for mass production and genetic transformation of KAT 60/8 variety.Key Words: 2, 4- D, embryogenic callus, shoot regeneration
Highlights
Pigeon pea (Cajanus cajan (L) Millsp.) is a drought tolerant pulse legume mainly grown for grain in the semi-arid tropics (Nene and Sheila, 1990; Pambo, 2014)
An initial seed germination of more than 95% was achieved with surface sterilised ICEAP 00557 and KAT 60/8 seeds at 4 days after inoculation, on PGRs-free MS medium, up to 100% germination was recorded after 7 days of culture (Table 1)
There were no bacterial or fungal contaminants observed after 10 days of culture; leaf explants obtained from the seedlings for subsequent experimentation were free from contamination
Summary
Pigeon pea (Cajanus cajan (L) Millsp.) is a drought tolerant pulse legume mainly grown for grain in the semi-arid tropics (Nene and Sheila, 1990; Pambo, 2014). It is an important food security crop for local consumption and export for several African countries, Kenya, Malawi, Mozambique, Tanzania and Uganda (Kaoneka et al, 2016). The main pigeon pea insect pest in Kenya is the pod borer (Helicoverpa armigera), which causes extensive damage to the crop (Lateef and Reed, 1983; Parde et al, 2012)
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