Establishment of In Vitro Regeneration System of Citrus australasica

doi:10.11983/CBB23060

Abstract

Abstract: In order to establish the regeneration system of Citrus australasica, the effects of different plant growth regulators combinations, medium types and dark culture time on callus induction and plant regeneration of C. australasica were studied using stem segments as explants. The results showed that the best medium for adventitious bud induction was 1/2MS+4.0 mg∙L^-1 ZT+30.0 g∙L^-1 sucrose. Dark culture for 14 days and then light culture had the best promoting effect. The induction rate of callus and adventitious bud was 100%, and the average number of adventitious bud regeneration per explants was 4.83. The optimal rooting medium was 1/2MS+0.5 mg∙L^-1 NAA, and the regenerated plants with the 94.43% rooting rate were obtained, and the average number of roots was 3.9. In the mixture of grass carbon:perlite: vermiculite=2:1:1 (v/v/v), tissue culture seedlings had the best growth, and the survival rate was more than 90%. This study established in vitro regeneration system of C. australasica, which laid the foundation for the genetic improvement and rapid propagation of C. australasica fine varieties.

Key words: Citrus australasica, stem segment, tissue culture, in vitro regeneration

Chungang Xie, Zhe Liu, Shusheng Zhang, Haitao Hu. Establishment of In Vitro Regeneration System of Citrus australasica[J]. Chinese Bulletin of Botany, 2023, 58(6): 926-934.

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URL: https://www.chinbullbotany.com/EN/10.11983/CBB23060

https://www.chinbullbotany.com/EN/Y2023/V58/I6/926

Figures/Tables 8

References 23

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No.	NAA (mg?L^-1)	6-BA (mg?L^-1)	3 weeks	5 weeks		Growth condition
No.	NAA (mg?L^-1)	6-BA (mg?L^-1)	Callus induction rate (%)	Rate of adventitious bud differentiation (%)	Frequency of bud differentiation	Growth condition
1	0	0	61.11	55.57±5.57 cde	1.57±0.29 bc	++
2	0	0.5	100.00	72.23±5.53 efg	2.08±0.08 de	++++
3	0	1.0	100.00	94.43±5.57 g	2.53±0.03 e	++++
4	0	2.0	91.67	55.57±5.57 cde	1.19±0.10 ab	++
5	0	4.0	58.33	27.73±5.57 abc	1.17±0.17 ab	+
6	0.1	0.5	58.33	61.13±5.57 def	2.36±0.07 de	+++
7	0.1	1.0	66.67	44.43±5.57 bcde	1.89±0.11 bce	++
8	0.1	2.0	75.00	44.47±11.17 bcde	2.00±0.29 bce	++
9	0.1	4.0	41.67	16.70±0.00 a	1.17±0.29 ab	+
10	0.5	0.5	66.67	55.57±5.57 cde	2.33±0.19 de	+++
11	0.5	1.0	91.67	66.67±9.61 ef	2.57±0.23 e	+++
12	0.5	2.0	33.33	33.33±9.61 abcd	1.61±0.20 bc	++
13	0.5	4.0	33.33	22.23±5.53 ab	1.17±0.17 ab	+

No.	NAA (mg?L^-1)	6-BA (mg?L^-1)	3 weeks	5 weeks		Growth condition
No.	NAA (mg?L^-1)	6-BA (mg?L^-1)	Callus induction rate (%)	Rate of adventitious bud differentiation (%)	Frequency of bud differentiation	Growth condition
1	0	0	61.11	55.57±5.57 cde	1.57±0.29 bc	++
2	0	0.5	100.00	72.23±5.53 efg	2.08±0.08 de	++++
3	0	1.0	100.00	94.43±5.57 g	2.53±0.03 e	++++
4	0	2.0	91.67	55.57±5.57 cde	1.19±0.10 ab	++
5	0	4.0	58.33	27.73±5.57 abc	1.17±0.17 ab	+
6	0.1	0.5	58.33	61.13±5.57 def	2.36±0.07 de	+++
7	0.1	1.0	66.67	44.43±5.57 bcde	1.89±0.11 bce	++
8	0.1	2.0	75.00	44.47±11.17 bcde	2.00±0.29 bce	++
9	0.1	4.0	41.67	16.70±0.00 a	1.17±0.29 ab	+
10	0.5	0.5	66.67	55.57±5.57 cde	2.33±0.19 de	+++
11	0.5	1.0	91.67	66.67±9.61 ef	2.57±0.23 e	+++
12	0.5	2.0	33.33	33.33±9.61 abcd	1.61±0.20 bc	++
13	0.5	4.0	33.33	22.23±5.53 ab	1.17±0.17 ab	+

No.	NAA (mg?L^-1)	ZT (mg?L^-1)	3 weeks	5 weeks		Growth condition
No.	NAA (mg?L^-1)	ZT (mg?L^-1)	Callus induction rate (%)	Rate of adventitious bud differentiation (%)	Frequency of bud differentiation	Growth condition
1	0	0	61.11	55.57±5.57 b	1.56±0.29 a	++
2	0	0.5	77.80	77.76±5.53 c	3.53±0.42 cdef	+++
3	0	1.0	83.30	88.87±5.57 c	3.63±0.75 cdef	++++
4	0	2.0	100.00	100.00±0.00 c	3.94±0.92 def	++++
5	0	3.0	100.00	100.00±0.00 c	3.94±0.53 defg	+++
6	0	4.0	100.00	100.00±0.00 c	4.43±0.50 fg	++++
7	0	5.0	100.00	100.00±0.00 c	4.24±0.20 fg	+++
8	0	6.0	83.33	77.77±5.53 c	3.19±0.19 bcdef	+++
9	0.1	0.5	88.90	77.76±5.53 c	2.50±0.30 abcde	++
10	0.1	1.0	100.00	100.00±0.00 c	3.55±0.48 cdef	++++
11	0.1	2.0	100.00	100.00±0.00 c	3.28±0.79 cdef	+++
12	0.1	3.0	100.00	100.00±0.00 c	3.28±0.46 bcdef	+++
13	0.1	4.0	94.44	94.43±5.57 c	4.21±0.45 efg	++++
14	0.1	5.0	94.44	88.87±5.57 c	3.42±0.14 cdef	+++
15	0.1	6.0	66.67	61.10±5.55 b	2.69±0.26 abd	+++
16	0.5	0.5	100.00	72.20±11.10 bc	1.49±0.16 ab	++
17	0.5	1.0	85.70	33.33±8.33 a	1.50±0.50 ab	++
18	0.5	2.0	88.90	50.00±9.64 ab	1.83±0.29 abc	++
19	0.5	3.0	88.89	50.00±9.64 ab	1.83±0.17 ab	++
20	0.5	4.0	83.33	88.87±5.57 c	2.86±0.17 abcde	+++
21	0.5	5.0	50.00	44.43±5.57 ab	2.27±0.37 abc	++
22	0.5	6.0	33.33	27.77±5.53 a	2.11±0.11 abc	+

No.	NAA (mg?L^-1)	ZT (mg?L^-1)	3 weeks	5 weeks		Growth condition
No.	NAA (mg?L^-1)	ZT (mg?L^-1)	Callus induction rate (%)	Rate of adventitious bud differentiation (%)	Frequency of bud differentiation	Growth condition
1	0	0	61.11	55.57±5.57 b	1.56±0.29 a	++
2	0	0.5	77.80	77.76±5.53 c	3.53±0.42 cdef	+++
3	0	1.0	83.30	88.87±5.57 c	3.63±0.75 cdef	++++
4	0	2.0	100.00	100.00±0.00 c	3.94±0.92 def	++++
5	0	3.0	100.00	100.00±0.00 c	3.94±0.53 defg	+++
6	0	4.0	100.00	100.00±0.00 c	4.43±0.50 fg	++++
7	0	5.0	100.00	100.00±0.00 c	4.24±0.20 fg	+++
8	0	6.0	83.33	77.77±5.53 c	3.19±0.19 bcdef	+++
9	0.1	0.5	88.90	77.76±5.53 c	2.50±0.30 abcde	++
10	0.1	1.0	100.00	100.00±0.00 c	3.55±0.48 cdef	++++
11	0.1	2.0	100.00	100.00±0.00 c	3.28±0.79 cdef	+++
12	0.1	3.0	100.00	100.00±0.00 c	3.28±0.46 bcdef	+++
13	0.1	4.0	94.44	94.43±5.57 c	4.21±0.45 efg	++++
14	0.1	5.0	94.44	88.87±5.57 c	3.42±0.14 cdef	+++
15	0.1	6.0	66.67	61.10±5.55 b	2.69±0.26 abd	+++
16	0.5	0.5	100.00	72.20±11.10 bc	1.49±0.16 ab	++
17	0.5	1.0	85.70	33.33±8.33 a	1.50±0.50 ab	++
18	0.5	2.0	88.90	50.00±9.64 ab	1.83±0.29 abc	++
19	0.5	3.0	88.89	50.00±9.64 ab	1.83±0.17 ab	++
20	0.5	4.0	83.33	88.87±5.57 c	2.86±0.17 abcde	+++
21	0.5	5.0	50.00	44.43±5.57 ab	2.27±0.37 abc	++
22	0.5	6.0	33.33	27.77±5.53 a	2.11±0.11 abc	+

No.	Different media	3 weeks	5 weeks		Growth condition
No.	Different media	Callus induction rate (%)	Rate of adventitious bud differentiation (%)	Frequency of bud differentiation	Growth condition
1	1/2MS	100.00	100.00±0.00 a	4.57±0.33 a	++++
2	MT	83.30	61.13±5.57 b	3.57±0.24 b	+++
3	MS	100.00	88.87±5.57 ab	3.64±0.38 b	+++
4	WPM	83.30	72.23±5.53 bc	2.83±0.17 bc	+++
5	White	66.70	33.33±0.00 e	1.56±0.29 c	+