Establishment of Immature Embryo Rescue and Regeneration System for Pyrus calleryana cv. ‘Cleveland’

doi:10.11983/CBB23152

Abstract

Abstract: In this study, the seed embryos of Pyrus calleryana cv. ‘Cleveland’ were used as materials to systematically study the sterilization of seed embryo explants, callus induction, adventitious bud proliferation and rooting medium. The results showed that the germination rate of young embryos stored at 4°C for 21 d was the highest (67.23%) The suitable sterilization treatment of seed embryo explants was 75% alcohol (30 s)+10% H₂O₂(10 min)+0.1% HgCl₂ (14 min)_.The most suitable medium for seed embryo germination was 1/2MS+4.0 mg·L^-1 6-BA+0.5 mg·L^-1 NAA, and the germination rate was 89.67%. The optimum culture medium for callus induction was 1/2MS+1.0 mg·L^-1 IBA+1.0 mg·L^-1 6-BA, and the callus induction rate was 93.33%. The best differentiation medium was 1/2MS+0.2 mg·L^-1 IBA+2.0 mg·L^-1 6-BA, and the regeneration frequency was 87.44%. Best transgenerational proliferation medium was 1/2MS+1.5 mg·L^-1 6-BA+0.1 mg·L^-1 IBA, and the proliferation rate was 100%. The most suitable medium for root formation was 1/2MS+ 20 g·L^-1 sucrose+1.0 g·L^-1 active carbon+1.5 mg·L^-1 IBA+0.05 mg·L^-1 NAA, and the rooting rate was 82.63%. In summary, the seed embryo regeneration system of P. calleryang cv. ‘Cleveland’ was established, which provides scientific basis and guidan-ce for the efficient reproduction of high-quality pear germplasm resources.

Key words: Pyrus calleryana cv. ‘Cleveland’, embryo rescue, tissue culture, regeneration system

Yuze Liu, Yifei Wang, Weizhen Ren, Hao Li, Bin Lu, Bingshe Lu, Xiaoyue Yu. Establishment of Immature Embryo Rescue and Regeneration System for Pyrus calleryana cv. ‘Cleveland’[J]. Chinese Bulletin of Botany, 2024, 59(5): 800-809.

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URL: https://www.chinbullbotany.com/EN/10.11983/CBB23152

https://www.chinbullbotany.com/EN/Y2024/V59/I5/800

Figures/Tables 10

References 31

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No.	6-BA (mg·L^-1)	IBA (mg·L^-1)
C1	1.0	0.05
C2	1.0	0.10
C3	1.0	0.20
C4	1.5	0.05
C5	1.5	0.10
C6	1.5	0.20
C7	2.0	0.05
C8	2.0	0.10
C9	2.0	0.20
C10	3.0	0.05
C11	3.0	0.10
C12	3.0	0.20
C13	4.0	0.05
C14	4.0	0.10
C15	4.0	0.20

No.	6-BA (mg·L^-1)	IBA (mg·L^-1)
C1	1.0	0.05
C2	1.0	0.10
C3	1.0	0.20
C4	1.5	0.05
C5	1.5	0.10
C6	1.5	0.20
C7	2.0	0.05
C8	2.0	0.10
C9	2.0	0.20
C10	3.0	0.05
C11	3.0	0.10
C12	3.0	0.20
C13	4.0	0.05
C14	4.0	0.10
C15	4.0	0.20

No.	Callus rate (%)	Differentiation rate (%)	Number of adventitious buds regenerated	Callus form
B1	60.56±2.55 e	77.22±2.55 c	3.27±0.04 b	Yellowish green, white, sticky lump
B2	66.11±2.55 d	72.78±2.55 d	2.79±0.04 c	Yellowish green, white, loose
B3	74.44±2.55 c	70.56±1.92 d	2.86±0.03 c	Light green, white, ropy
B4	72.22±2.55 c	89.44±1.92 a	3.79±0.06 a	Light green, loose
B5	85.56±1.92 b	82.78±1.92 b	3.86±0.03 a	Chartreuse, white, ropy
B6	93.33±1.67 a	63.89±1.92 e	2.67±0.03 d	Chartreuse, white, ropy
B7	82.78±2.55 b	81.67±2.89 b	3.59±0.04 b	Yellow, white, clumpy and compact
B8	86.67±3.33 b	60.56±1.92 e	2.69±0.05 d	Light yellow, granular sphericity, compact and fragile
B9	91.67±2.89 a	61.67±1.67 e	2.38±0.04 e	Light yellow, white, waterlogged callus

No.	Callus rate (%)	Differentiation rate (%)	Number of adventitious buds regenerated	Callus form
B1	60.56±2.55 e	77.22±2.55 c	3.27±0.04 b	Yellowish green, white, sticky lump
B2	66.11±2.55 d	72.78±2.55 d	2.79±0.04 c	Yellowish green, white, loose
B3	74.44±2.55 c	70.56±1.92 d	2.86±0.03 c	Light green, white, ropy
B4	72.22±2.55 c	89.44±1.92 a	3.79±0.06 a	Light green, loose
B5	85.56±1.92 b	82.78±1.92 b	3.86±0.03 a	Chartreuse, white, ropy
B6	93.33±1.67 a	63.89±1.92 e	2.67±0.03 d	Chartreuse, white, ropy
B7	82.78±2.55 b	81.67±2.89 b	3.59±0.04 b	Yellow, white, clumpy and compact
B8	86.67±3.33 b	60.56±1.92 e	2.69±0.05 d	Light yellow, granular sphericity, compact and fragile
B9	91.67±2.89 a	61.67±1.67 e	2.38±0.04 e	Light yellow, white, waterlogged callus

No.	6-BA (mg·L^-1)	IBA (mg·L^-1)	Multiplication rates (%)	Multiplication coefficient	Growth condition
C1	1.0	0.05	91.11±0.96 d	3.17±0.17 efg	Average, short
C2	1.0	0.1	87.22±0.96 e	3.00±0.44 fg	Average, short
C3	1.0	0.2	95.00±1.67 c	2.89±0.10 g	Average
C4	1.5	0.05	100.00±0.00 a	4.17±0.44 bc	Stronger
C5	1.5	0.1	100.00±0.00 a	5.22±0.35 a	Stronger, large leaf blade, dark green
C6	1.5	0.2	100.00±0.00 a	4.11±0.10 bcd	Stronger, dark green
C7	2.0	0.05	100.00±0.00 a	3.67±0.17 cde	Stronger, dark green
C8	2.0	0.1	100.00±0.00 a	4.50±0.33 b	Stronger
C9	2.0	0.2	100.00±0.00 a	3.61±0.19 cdef	Stronger
C10	3.0	0.05	95.56±0.96 c	3.61±0.69 cdef	Average growth, small leaf blade
C11	3.0	0.1	97.22±0.96 b	3.50±0.44 defg	Average growth, small leaf blade
C12	3.0	0.2	100.00±0.00 a	3.28±0.19 efg	Average growth, dwarf tufted bud, deformed seedling
C13	4.0	0.05	96.11±0.96 bc	3.94±0.35 bcd	Weak growth, fewer leaves, red stems and leaves, glass seedlings
C14	4.0	0.1	92.22±0.96 d	3.67±0.29 cde	Small leaf blade, dwarf tufted bud, glass seedlings, deformed seedling
C15	4.0	0.2	100.00±0.00 a	3.56±0.19 cdef	Malformed leaf, glass seedlings