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First published online 1 October 2003
doi: 10.1242/dev.00794

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Genetic analysis of adventitious root formation with a novel series of temperature-sensitive mutants of Arabidopsis thaliana

Botanical Gardens, Graduate School of Science, The University of Tokyo, Hakusan 3-7-1, Bunkyo-ku, Tokyo 112-0001, Japan

View larger version (57K): [in a new window]   Fig. 1. Adventitious root induction from hypocotyl explants cultured on RIM for 16 days at 22°C (A-K) or at 28°C (L-V). (A,L) Wild type, (B,M) srd2, (C,N) rid1, (D,O) rid2, (E,P) rid3, (F,Q) rid4, (G,R) rid5, (H,S) rpd1, (I,T) rgd1, (J,U) rgd2, (K,V) rgd3. Scale bar: 5 mm.

View larger version (158K): [in a new window]   Fig. 2. Early stage of adventitious root formation. Hypocotyl explants were cultured on RIM at 22°C (A,C,E,G,I,K,M,O,Q,S,U) or at 28°C (B,D,F,H,J,L,N,P,R,T,V). (A,B) Wild type at day 6, (C,D) srd2 at day 11, (E,F) rid1 at day 16, (G,H) rid2 at day 6, (I,J) rid3 at day 6, (K,L) rid4 at day 6, (M,N) rid5 at day 6, (O,P) rpd1 at day 6, (Q,R) rgd1 at day 6, (S,T) rgd2 at day 6, (U,V) rgd3 at day 6. Scale bar: 100 µm.

View larger version (19K): [in a new window]   Fig. 3. Effect of auxin concentration on the rooting rate of the rid5 mutant. Hypocotyl explants of the wild type and rid5 were cultured on RIM containing various concentrations of IBA at either 22°C or 28°C. Each symbol represents an average value of data obtained from two independent experiments, in which 25 hypocotyls were used for every point. Vertical lines indicate standard deviation.

View larger version (98K): [in a new window]   Fig. 4. Lateral root formation. Primary root explants were cultured on RIM for 16 days at 22°C (A,C,E,G) or 28°C (B,D,F,H). (A, B) Wild type, (C,D) srd2, (E,F) rid1, (G,H) rid2. Scale bar: 100 µm.

View larger version (49K): [in a new window]   Fig. 5. Callus formation from hypocotyl explants. Hypocotyl explants were cultured on CIM for 24 days at 22°C (A-K) or at 28°C (L-V). (A,L) Wild type, (B,M) srd2, (C,N) rid1, (D,O) rid2, (E,P) rid3, (F,Q) rid4, (G,R) rid5, (H,S) rpd1, (I,T) rgd1, (J,U) rgd2, (K,V) rgd3. Scale bar: 5 mm.

View larger version (58K): [in a new window]   Fig. 6. Callus formation from root explants. Root explants were cultured on CIM for 24 days at 22°C (A–K,W,X) or at 28°C (L-V,Y,Z). (A,L) Wild type, (B,M) srd2, (C,N) rid1, (D,O) rid2, (E,P) rid3, (F,Q) rid4, (G,R) rid5, (H,S) rpd1, (I,T) rgd1, (J,U) rgd2, (K,V) rgd3. (W-Z) Higher magnification of the wild type (W,Y) and rid2 (X,Z). Scale bar: 5 mm.

View larger version (119K): [in a new window]   Fig. 7. Phenotypes of seedlings that were grown for 12 days on vertical GMA plates. (A) Seedlings grown at 22°C. (B) Seedlings grown at 28°C. (C-J) Epidermis of the hypocotyl (C-F) and primary root (G-J) of the wild type and rid5. (C,G) Wild type at 22°C, and (D,H) at 28°C. (E,I) rid5 at 22°C. (F,J) rid5 at 28°C. Arrows indicate the direction of growth. Epidermal cell files of the rid5 seedling showed left-handed helical growth at 28°C. Arrowheads in (J) indicate branched root hairs. (K-M) True leaves of the wild type (K) and rid3 (L,M) at 28°C. Bars in (A,B), 1 cm. Scale bars: (C-J) 200 µm; (K,L,M), 500 µm.

View larger version (28K): [in a new window]   Fig. 8. A hypothetical scheme for adventitious root formation.