Chin Bull Bot ›› 2019, Vol. 54 ›› Issue (5): 634-641.doi: 10.11983/CBB18193

• TECHNIQUES AND METHODS • Previous Articles     Next Articles

The Effect of Different Hormones on Explant Browning and Callus Browning in Cyclocarya paliurus

Feng Ying1,*(),Qian Lianwen1,Lin Qingliang2   

  1. 1. School of Resource and Environmental Science, Quanzhou Normal University, Quanzhou 362000, China
    2. Fujian Agriculture and Forestry University, Fuzhou 350002, China
  • Received:2018-09-11 Accepted:2019-02-19 Online:2020-03-10 Published:2019-09-01
  • Contact: Feng Ying E-mail:fengy0919@126.com

Abstract:

To determine the effect of different hormones on explant browning and callus browning in Cyclocarya paliurus, leaf browning was controlled on medium with different kinds and content of hormones (6-BA, GA3, NAA) by a single-factor completely random test. Callus browning was controlled on the medium with different content of 6-BA+NAA and different basic medium by a double- and single-factor completely random test, respectively. The rate of leaf browning reached 0, and the rate of callus induction was up to 100% on the improved MS medium with 1.0 mg∙L -1 6-BA and 1.0 mg∙L -1 GA3 and 0.3 mg∙L -1 NAA. The rate of callus browning reached 0, and the callus propagation was up to 4.80 times on the improved MS medium with 0.5 mg∙L -1 6-BA and 0.2 mg∙L -1NAA. The callus was yellow with green or yellow, granular, small and compact, hard and dry. This system effectively solved the problem of leaf browning and callus browning and provided a simple and effective way for tissue culture and a solid foundation for the regeneration system from leaf of C. paliurus.

Key words: browning, Cyclocarya paliurus, callus, induction, propagation

Table 1

6-BA+GA3+NAA hormone combination"

Treatment 6-BA content (mg·L-1) GA3 content (mg·L-1) NAA content (mg·L-1)
6-BA1 1.0 1.0 0.3
6-BA2 2.0 1.0 0.3
6-BA3 3.0 1.0 0.3
GA31 1.0 0.0 0.3
GA32 1.0 0.5 0.3
GA33 1.0 1.0 0.3
GA34 1.0 1.5 0.3
NAA1 1.0 1.0 0.0
NAA2 1.0 1.0 0.1
NAA3 1.0 1.0 0.3
NAA4 1.0 1.0 0.5

Figure 1

The type of callus induced from leaves of Cyclocarya paliurus (A) Callus of the type I; (B) Callus of the type II; (C) Callus of the type III. Bars=1 cm"

Table 2

The effect of 6-BA on leaf browning in Cyclocarya paliurus (means±SD)"

Medium The rate of callus induction (%) The rate of leaf browning (%) Callus
Type Growth
1.0 mg·L-1 6-BA+1.0 mg·L-1 GA3+0.3 mg·L-1 NAA 100.00±0.00 a 0.00±0.00 a I, II I (++), II (+)
2.0 mg·L-1 6-BA+1.0 mg·L-1 GA3+0.3 mg·L-1 NAA 95.24±8.25 a 4.76±8.25 b I, II I (+++), II (++)
3.0 mg·L-1 6-BA+1.0 mg·L-1 GA3+0.3 mg·L-1 NAA 66.67±18.04 b 33.33±18.04 b I, III I (++), III (+)

Table 3

The effect of GA3 on leaf browning in Cyclocarya paliurus (means±SD)"

Medium The rate of callus induction (%) The rate of leaf browning (%) Callus
Type Growth
1.0 mg·L-1 6-BA+0.3 mg·L-1 NAA 48.96±4.77 c 51.04±4.77 a I, III I (++), III (+)
0.5 mg·L-1 GA3+1.0 mg·L-1 6-BA+0.3 mg·L-1 NAA 86.67±10.41 b 13.33±10.41 b I, II, III I (++), II (+), III (-)
1.0 mg·L-1 GA3+1.0 mg·L-1 6-BA+0.3 mg·L-1 NAA 100.00±0.00 a 0.00±0.00 c I, II I (++), II (+)
1.5 mg·L-1 GA3+1.0 mg·L-1 6-BA+0.3 mg·L-1 NAA 86.11±2.78 b 13.89±2.78 b I I (++)

Figure 2

The effect of GA3 on callus induction of leaves in Cyclocarya paliurus (A) Callus in the induction medium without GA3; (B) Callus cultured for 15 days in the induction medium with GA3; (C) Callus cultured for 30 days in the induction medium with GA3. Bars=1 cm"

Table 4

The effect of NAA on leaf browning in Cyclocarya paliurus (means±SD)"

Medium The rate of callus induction (%) The rate of leaf browning (%) Callus
Type Growth
1.0 mg·L-1 GA3+1.0 mg·L-1 6-BA 23.15±11.25 c 76.85±11.25 a I I (-)
0.1 mg·L-1 NAA+1.0 mg·L-1 GA3+1.0 mg·L-1 6-BA 71.39±7.01 b 18.61±7.01 b I, II I (++), II (+)
0.3 mg·L-1 NAA+1.0 mg·L-1 GA3+1.0 mg·L-1 6-BA 100.00±0.00 a 0.00±0.00 c I, II I (++), II (+)
0.5 mg·L-1 NAA+1.0 mg·L-1 GA3+1.0 mg·L-1 6-BA 96.99±0.20 a 3.11±0.20 c I, II I (+), II (+)

Figure 3

The type of callus propagation in Cyclocarya paliurus (A), (B) The callus of type I; (C), (D) The callus of type II; (E), (F) The callus of type III. Bars=1 cm"

Table 5

The effect of 6-BA+NAA on callus browning in Cyclocarya paliurus"

Plant growth regulator (mg·L-1) The time of callus propagation (time) The rate of callus browning (%) Callus
6-BA NAA Type Growth
0.2 0.05 1.25 de 0 e II II (+++)
0.5 0.05 3.87 b 0 e I, II I (+++), II (++)
0.8 0.05 3.51 b 0 e I, II I (++), II (+++)
1.0 0.05 0.69 e 0 e I, III I (++), III (+)
0.2 0.2 0.75 e 0 e I, II, III I (+), II (+), III (+)
0.5 0.2 4.80 a 0 e I, II I (++), II (+++)
0.8 0.2 3.55 b 0 e I, II, III I (++), II (+++), III (++++)
1.0 0.2 1.30 de 40.00 a I, II I (++), II (+)
0.2 0.4 0.97 de 33.33 ab I, III I (+), III (+)
0.5 0.4 1.65 d 28.33 bc II, III II (+), III (+)
0.8 0.4 2.57 c 20.00 d II, III II (++), III (+)
1.0 0.4 1.75 d 21.67 cd II, III II (+), III (+)

Table 6

The effect of 6-BA and NAA on callus browning in Cyclocarya paliurus, respectively (means±SD)"

Plant growth regulator The time of callus propagation (time) The rate of callus browning (%)
0.2 mg·L-1 6-BA 0.98±0.33 b 11.11±17.64 b
0.5 mg·L-1 6-BA 3.44±1.47 a 9.44±15.09 b
0.8 mg·L-1 6-BA 3.21±0.67 a 6.67±10.00 b
1.0 mg·L-1 6-BA 1.25±0.55 b 20.56±17.40 a
0.05 mg·L-1 NAA 2.33±1.49 a 0.00±0.00 c
0.2 mg·L-1 NAA 2.59±1.75 a 10.00±18.09 b
0.4 mg·L-1 NAA 1.73±0.74 b 25.83±8.75 a

Figure 4

The effect of basic medium on callus browning (A) and callus propagation (B) in Cyclocarya paliurus (means±SD) Different lowercase letters indicate significant differences at P<0.05."

Figure 5

Callus of Cyclocarya paliurus growth in the different basic medium (A) Callus growth in the improved MS basic medium; (B), (C) Callus growth in the improved MS1 basic medium; (D), (E) Callus growth in the improved MS2 basic medium; (F) Callus growth in the improved MS3 basic medium. Bars=1 cm"

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