Establishment of Callus Regeneration System of Hippeastrum ‘Bangkok Rose’

doi:10.3969/j.issn.1000-2561.2023.05.013

Abstract

Abstract:

Hippeastrum is a new kind of flower introduced in large scale from abroad in recent years, but its seedlings are expensive, regular scale cuttage propagation speed is slow and needs a large number of mother bulbs. Cross breeding, a common method for its new varieties, however, has poor orientation. This aim of this study was to establish an efficient callus regeneration system for rapid propagation and factory production of seedlings, which can also be used for the orientation breeding of Hippeastrum. The effects about different plant growth regulators on callus induction and plantlet regeneration were studied with the leaves of Hippeastrum ‘Bankkok Rose’ plantlet in vitro. The highest induction rate of callus (39.67%) was observed when the basal leaves of the buds in vitro were cultured on MS+2,4-D 2.00 mg/L+ TDZ 0.50 mg/L medium for 45 days. The optimal medium for callus proliferation was MS+6-BA 2.00 mg/L, and the average proliferation was 4.01 times every 20 days. The optimal medium for callus differentiation was MS+ KT 0.50 mg/L. After 60 days, the adventitious bud differentiation coefficient reached 10.59, and the seedling formation coefficient was 5.67. The rooting rate reached 100% after 30 days in MS+IBA 0.50 mg/L. After 30 days of root culture, the small plants were transplanted to the coir: peat soil: vermiculite =1 : 1 : 1 substrate, and the survival rate reached 93.33% after 30 days. This study could provide technical support for industrial propagation of Hippeastrum seedlings, and also provide excellent receptor materials for subsequent molecular breeding.

Key words: Hippeastrum, leave in vitro, plant growth regulators, tissue culture

CLC Number:

S682.31

YANG Wei, LIU Xinyi, ZENG Jingjue, WU Kuilin, FANG Lin, WU Shasha, ZHAI Junwen, ZENG Songjun. Establishment of Callus Regeneration System of Hippeastrum ‘Bangkok Rose’[J]. Chinese Journal of Tropical Crops, 2023, 44(5): 977-985.

Figures/Tables 7

References 36

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6-BA浓度 Concentration of 6-BA/(mg·L^-1)	生物增重倍数 Biological weight gain multiple	愈伤组织增重倍数 Callus weight gain multiple	不定芽分化率 Adventitious bud differentiation rate/%	生长状态 Growth status
0.00	3.86±0.21^b	1.50±0.14^c	100.00±0.00^a	芽点较多，淡绿色，质地较脆
1.00	5.65±0.58^a	3.21±0.66^ab	50.00±5.77^b	下部橙黄色，部分有芽点
2.00	4.01±0.54^b	4.01±0.53^a	0.00±0.00^c	鲜黄色，质地蓬松
4.00	2.96±0.04^b	2.96±0.07^b	0.00±0.00^c	基部稍褐化，鲜黄色
6.00	2.84±0.24^b	2.84±0.24^b	0.00±0.00^c	基部褐化变黑，呈褐黄色

6-BA浓度 Concentration of 6-BA/(mg·L^-1)	生物增重倍数 Biological weight gain multiple	愈伤组织增重倍数 Callus weight gain multiple	不定芽分化率 Adventitious bud differentiation rate/%	生长状态 Growth status
0.00	3.86±0.21^b	1.50±0.14^c	100.00±0.00^a	芽点较多，淡绿色，质地较脆
1.00	5.65±0.58^a	3.21±0.66^ab	50.00±5.77^b	下部橙黄色，部分有芽点
2.00	4.01±0.54^b	4.01±0.53^a	0.00±0.00^c	鲜黄色，质地蓬松
4.00	2.96±0.04^b	2.96±0.07^b	0.00±0.00^c	基部稍褐化，鲜黄色
6.00	2.84±0.24^b	2.84±0.24^b	0.00±0.00^c	基部褐化变黑，呈褐黄色

6-BA浓度Concentration of 6-BA/(mg·L^-1)	KT浓度 Concentration of KT/(mg·L^-1)	NAA浓度 Concentration of NAA/(mg·L^-1)	分化率 Differentiation rate/%	分化系数 Differentiation coefficient	平均每团愈伤组织出苗数 Average number of seedlings produced from each callus group	平均株高 Average plant height/cm
0.25	0.00	0.00	83.33±8.33^ab	3.62±0.72^ghl	0.50±0.25^fg	3.83±0.44^bcd
0.25	0.00	0.50	58.33±8.33^b	3.04±0.40^l	1.17±0.83^defg	3.17±0.44^cde
0.50	0.00	0.00	83.33±16.67^ab	4.34±0.34^gh	1.83±0.41^cdef	4.33±0.33^bcd
0.50	0.00	0.50	58.33±22.05^b	3.00±0.40^l	0.75±0.25^defg	3.67±0.44^bcd
0.75	0.00	0.00	91.67±8.33^a	3.96±0.42^ghl	0.25±0.14^g	3.50±0.50^bcd
0.75	0.00	0.50	83.33±8.33^ab	2.75±0.14^l	0.58±0.22^efg	5.83±0.73^ab
1.00	0.00	0.00	58.33±16.67^b	3.96±0.11^ghl	0.58±0.22^efg	4.00±1.00^bcd
1.00	0.00	0.10	76.67±12.01^ab	3.72±0.26^ghl	1.07±0.97^defg	1.83±0.93^de
1.00	0.00	0.20	73.33±3.33^ab	3.10±0.60^hl	0.20±0.57^g	2.33±0.17^cde
1.00	0.00	0.50	58.33±8.33^b	2.92±0.50^l	0.58±0.22^efg	2.67±0.17^cde
1.00	0.00	1.00	58.33±4.41^b	1.30±0.12ⁱ	0.17±0.67^g	2.07±0.67^de
0.00	0.25	0.00	100.00±0.00^a	7.88±0.31^bc	3.42±0.22^b	4.33±0.17^bcd
0.00	0.25	0.50	91.67±8.33^a	6.92±0.36^cd	1.42±0.22^cdefg	4.00±0.58^bcd
0.00	0.50	0.00	100.00±0.00^a	10.59±0.18^a	5.67±1.01^a	6.83±0.73^a
0.00	0.50	0.50	91.67±8.33^a	7.84±0.29^bc	0.92±0.68^defg	2.33±1.20^cde
0.00	0.75	0.00	100.00±0.00^a	8.33±0.300^b	2.58±0.30^bc	4.83±0.44^abc
0.00	0.75	0.50	83.33±8.33^ab	5.63±0.94^ef	0.25±0.25^g	1.00±1.00^e
0.00	1.00	0.00	100.00±0.00^a	8.04±0.42^bc	2.00±0.25^cde	4.33±1.33^bcd
0.00	1.00	0.50	91.67±8.33^a	4.63±0.07^fg	0.25±0.14^g	3.17±1.59^cde
0.00	0.00	0.00	100.00±0.00^a	5.88±0.50^de	2.08±0.58^bcd	4.00±0.50^bcd

6-BA浓度Concentration of 6-BA/(mg·L^-1)	KT浓度 Concentration of KT/(mg·L^-1)	NAA浓度 Concentration of NAA/(mg·L^-1)	分化率 Differentiation rate/%	分化系数 Differentiation coefficient	平均每团愈伤组织出苗数 Average number of seedlings produced from each callus group	平均株高 Average plant height/cm
0.25	0.00	0.00	83.33±8.33^ab	3.62±0.72^ghl	0.50±0.25^fg	3.83±0.44^bcd
0.25	0.00	0.50	58.33±8.33^b	3.04±0.40^l	1.17±0.83^defg	3.17±0.44^cde
0.50	0.00	0.00	83.33±16.67^ab	4.34±0.34^gh	1.83±0.41^cdef	4.33±0.33^bcd
0.50	0.00	0.50	58.33±22.05^b	3.00±0.40^l	0.75±0.25^defg	3.67±0.44^bcd
0.75	0.00	0.00	91.67±8.33^a	3.96±0.42^ghl	0.25±0.14^g	3.50±0.50^bcd
0.75	0.00	0.50	83.33±8.33^ab	2.75±0.14^l	0.58±0.22^efg	5.83±0.73^ab
1.00	0.00	0.00	58.33±16.67^b	3.96±0.11^ghl	0.58±0.22^efg	4.00±1.00^bcd
1.00	0.00	0.10	76.67±12.01^ab	3.72±0.26^ghl	1.07±0.97^defg	1.83±0.93^de
1.00	0.00	0.20	73.33±3.33^ab	3.10±0.60^hl	0.20±0.57^g	2.33±0.17^cde
1.00	0.00	0.50	58.33±8.33^b	2.92±0.50^l	0.58±0.22^efg	2.67±0.17^cde
1.00	0.00	1.00	58.33±4.41^b	1.30±0.12ⁱ	0.17±0.67^g	2.07±0.67^de
0.00	0.25	0.00	100.00±0.00^a	7.88±0.31^bc	3.42±0.22^b	4.33±0.17^bcd
0.00	0.25	0.50	91.67±8.33^a	6.92±0.36^cd	1.42±0.22^cdefg	4.00±0.58^bcd
0.00	0.50	0.00	100.00±0.00^a	10.59±0.18^a	5.67±1.01^a	6.83±0.73^a
0.00	0.50	0.50	91.67±8.33^a	7.84±0.29^bc	0.92±0.68^defg	2.33±1.20^cde
0.00	0.75	0.00	100.00±0.00^a	8.33±0.300^b	2.58±0.30^bc	4.83±0.44^abc
0.00	0.75	0.50	83.33±8.33^ab	5.63±0.94^ef	0.25±0.25^g	1.00±1.00^e
0.00	1.00	0.00	100.00±0.00^a	8.04±0.42^bc	2.00±0.25^cde	4.33±1.33^bcd
0.00	1.00	0.50	91.67±8.33^a	4.63±0.07^fg	0.25±0.14^g	3.17±1.59^cde
0.00	0.00	0.00	100.00±0.00^a	5.88±0.50^de	2.08±0.58^bcd	4.00±0.50^bcd

IBA浓度 Concentration of IBA /(mg·L^-1)	NAA浓度 Concentration of NAA /(mg·L^-1)	生根率 Rooting rate/%	平均根数 Average root number	平均根长 Average root length/cm	平均株高 Average plant height/cm
0.00	0.00	100±0.00^a	4.30±0.42^bc	2.63±0.24^a	3.89±0.13^b
0.50	0.00	100±0.00^a	4.85±0.26^abc	1.48±0.19^c	6.00±0.35^a
1.00	0.00	95.00±5.00^ab	4.85±0.5^abc	0.85±0.12^def	4.75±0.43^ab
1.50	0.00	80.00±8.16^abc	4.50±0.71^bc	0.55±0.09^f	4.50±0.20^ab
0.00	0.10	95.00±5.00^ab	4.65±0.54^bc	2.00±0.20^b	5.50±0.35^ab
0.50	0.10	95.00±5.00^ab	4.45±0.49^bc	0.83±0.26^def	4.88±0.38^ab
1.00	0.10	100.00±0.00^a	6.57±0.49^a	1.28±0.75^cd	4.38±0.75^ab
1.50	0.10	70.00±12.91^c	3.25±0.78^c	0.48±0.25^f	5.38±0.66^ab
0.00	0.20	80.00±11.55^abc	3.60±0.63^c	1.23±0.15^cde	4.75±0.32^ab
0.50	0.20	95.00±5.00^ab	5.65±0.33^ab	1.23±0.10^cde	5.37±0.43^ab
1.00	0.20	70.00±10.00^c	4.00±0.38^bc	0.78±0.11^ef	5.13±0.66^ab
1.50	0.20	75.00±5.00^bc	5.05±0.38^abc	0.85±0.65^def	5.45±0.89^ab