Optimization of Callus Induction from Immature Embryo and Establishment Regeneration System of Paeonia ostii‘Fengdan’

doi:10.16420/j.issn.0513-353x.2020-0243

Acta Horticulturae Sinica ›› 2022, Vol. 49 ›› Issue (1): 166-174.doi: 10.16420/j.issn.0513-353x.2020-0243

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Optimization of Callus Induction from Immature Embryo and Establishment Regeneration System of Paeonia ostii‘Fengdan’

LIU Rong, CI Huiting, REN Xiuxia, GAO Jie, WANG Shunli(), ZHANG Xiuxin()

Key Laboratory of Biology and Genetic Improvement of Horticultural Crops,Ministry of Agriculture,P. R. China,Research Center of Peony,Chinese Academy of Agricultural Sciences,National Center for Flower Improvement,Institute of Vegetables and Flowers,Chinese Academy of Agricultural Sciences,Beijing 100081,China

Received:2021-10-20 Revised:2021-11-18 Online:2022-01-25 Published:2022-01-24

Abstract

Abstract:

In order to improve callus induction quality,and establish highly efficient regeneration system of Paeonia ostii‘Fengdan’,the immature embryo collecting time,technical treatments of immature embryos,basic medium selection,and different concentrations and combinations of plant growth regulators and H₂O₂ were optimized. The results showed that the best collection time for immature embryos callus induction was 60 days after pollination. Compared with four technical treatments of immature embryos,thoroughly divided embryos（slices）were produced better callus with low callus browning rate than that with whole embryos,halved embryos,and broken into quarters embryos. MS basic medium with 0.162 μmol · L^-1 H₂O₂ can improve the callus induction rate of immature embryo,and it can also increases the callus browning rate. Different concentration of H₂O₂ adding in WPM basic medium could not be improved the callus induction rate. WPM basic medium is better than MS basic medium for immature embryo callus induction. The combination of WPM + 2.5 mg · L^-1 2,4-D + 0.1 mg · L^-1 6-BA can obtain higher callus induction rate（93.33% ± 3.53%）with lower callus browning rate（25.33% ± 4.81%）. The best hormone combinations for adventitious bud and root induction were WPM + 0.5 mg · L^-1 GA₃ + 0.5 mg · L^-1 6-BA,1/2 WPM + 2.0 mg · L^-1 IBA + 1.0 mg · L^-1 IAA,respectively.

Key words: tree peony, immature embryo, callus induction, callus browning, in vitro, regeneration

CLC Number:

S685.11

LIU Rong, CI Huiting, REN Xiuxia, GAO Jie, WANG Shunli, ZHANG Xiuxin. Optimization of Callus Induction from Immature Embryo and Establishment Regeneration System of Paeonia ostii‘Fengdan’[J]. Acta Horticulturae Sinica, 2022, 49(1): 166-174.

Figures/Tables 6

References 35

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取材时间 Embryo collection time	愈伤组织诱导率/% Callus induction rate	褐化率/% Browning rate
授粉后50 d 50 days after pollination	86.67 ± 2.81 B	4.00 ± 2.31 C
授粉后60 d 60 days after pollination	97.33 ± 2.67 A	4.00 ± 0.00 C
授粉后70 d 70 days after pollination	90.67 ± 2.67 C	21.33 ± 3.53 B
授粉后80 d 80 days after pollination	66.67 ± 3.53 D	58.67 ± 1.33 A

取材时间 Embryo collection time	愈伤组织诱导率/% Callus induction rate	褐化率/% Browning rate
授粉后50 d 50 days after pollination	86.67 ± 2.81 B	4.00 ± 2.31 C
授粉后60 d 60 days after pollination	97.33 ± 2.67 A	4.00 ± 0.00 C
授粉后70 d 70 days after pollination	90.67 ± 2.67 C	21.33 ± 3.53 B
授粉后80 d 80 days after pollination	66.67 ± 3.53 D	58.67 ± 1.33 A

接种方法 Inoculation method	愈伤诱导率/% Callus induction rate	褐化率/% Browning rate	愈伤组织特征 Characteristics of callus
完整胚The intact embryo	100.00	60.00 ± 1.67 a	白色,较松软 White,soft
1/2胚 The 1/2 embryo	100.00	45.00 ± 3.33 b	白色,表面有雪花和松散的结构 White,with snowflakes and loose structure on the surface
1/4胚 The 1/4 embryo	100.00	25.00 ± 1.67 c	淡黄色,结构紧密 Light yellow,compact structure
幼胚薄片 The young embryo slice	100.00	15.00 ± 1.67 d	淡黄色,结构紧密,颗粒状 Light yellow,compact structure,granular

接种方法 Inoculation method	愈伤诱导率/% Callus induction rate	褐化率/% Browning rate	愈伤组织特征 Characteristics of callus
完整胚The intact embryo	100.00	60.00 ± 1.67 a	白色,较松软 White,soft
1/2胚 The 1/2 embryo	100.00	45.00 ± 3.33 b	白色,表面有雪花和松散的结构 White,with snowflakes and loose structure on the surface
1/4胚 The 1/4 embryo	100.00	25.00 ± 1.67 c	淡黄色,结构紧密 Light yellow,compact structure
幼胚薄片 The young embryo slice	100.00	15.00 ± 1.67 d	淡黄色,结构紧密,颗粒状 Light yellow,compact structure,granular

培养基 Culture media	H₂O₂/（μmol · L^-1）	愈伤组织诱导率/% Callus induction rate	愈伤组织褐化率/% Browning rate of callus
MS	0（对照 Control）	66.67 ± 10.90 b	28.00 ± 2.31 b
	0.162	90.67 ± 3.53 a	41.33 ± 13.33 a
	0.324	84.00 ± 2.31 ab	49.33 ± 12.72 a
	0.486	72.00 ± 8.33 b	29.33 ± 2.31 b
WPM	0（对照 Control）	97.33 ± 2.67 a	4.00 ± 0.00 c
	0.162	97.33 ± 1.33 a	4.00 ± 2.31 c
	0.324	92.00 ± 4.00 a	2.67 ± 1.33 c
	0.486	92.00 ± 4.62 a	2.67 ± 2.67 c

Optimization of Callus Induction from Immature Embryo and Establishment Regeneration System of Paeonia ostii‘Fengdan’

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植物生长调节剂 Plant growth regulator	浓度/（mg · L^-1） Concentration	愈伤诱导率/% Callus induction rate	褐化率/% Browning rate of callus
对照Control	0	16.00 ± 2.31 f	6.67 ± 1.33 i
2,4-D	1.00	90.67 ± 5.81 ab	69.33 ± 6.11 a
	1.50	85.33 ± 4.81 abc	69.33 ± 5.81 a
	2.00	90.67 ± 2.67 ab	58.67 ± 4.81 ab
	2.50	93.33 ± 3.53 ab	25.33 ± 4.81 fg
	3.00	97.33 ± 2.67 a	30.67 ± 2.67 ef
毒莠定Picloram	2.00	82.67 ± 10.41 abcd	22.20 ± 4.81 cde
	3.00	73.33 ± 8.74 cd	34.67 ± 3.53 def
	4.00	97.33 ± 2.67 a	46.67 ± 9.61 cde
	5.00	97.33 ± 2.67 a	56.00 ± 10.07 ab
麦草畏Dicamba	1.22	46.67 ± 3.53 e	20.00 ± 4.62 gh
	1.72	66.67 ± 5.33 d	44.00 ± 4.00 cd
	2.22	73.33 ± 1.33 cd	53.33 ± 1.33bc
	2.72	77.33 ± 5.81 bcd	48.00 ± 9.24 cd
	3.22	78.67 ± 5.81 bcd	8.00 ± 1.33 h

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