Ex vitro Rooting, Acclimatization and Genetic Stability of Lonicera caerulea var. kamtschatica

Primers, polymorphism between tested cultivars and amplification products of ISSR-PCR and AFLPPCR used for evaluation of genetic identity of Lonicera caerulea ‘Wojtek’ and ‘Zojka’ plants

No.	Primer	Sequence (5′–3′)	Total number of bands	Polymorphic bands (%)	Primer	Sequence (5′–3′)	Total number of bands	Polymorphic bands (%)
	ISSR				AFLP

1	UBC 806	(TA)₈G	-	-	Pst-TT/Mse-CC^*	GACTGCGTACATGCAGTT GATGAGTCCTGAGTAACC	30	13.3
2	UBC 810^*	(GA)₈T	6	16.7	Pst-GC/Mse-GC	GACTGCGTACATGCAGGC GATGAGTCCTGAGTAAGC	21	0
3	UBC 813	(CT)₈T	-	-	Pst-GC/Mse-TA^*	GACTGCGTACATGCAGGC GATGAGTCCTGAGTAATA	18	11.1
4	UBC 821	(GT)₈T	-	-	Pst-TA/Mse-TA	GACTGCGTACATGCAGTA GATGAGTCCTGAGTAATA	14	0
5	UBC 822	(TC)₈A	9	0	Pst-TA/Mse-GC	GACTGCGTACATGCAGTA GATGAGTCCTGAGTAAGC	42	0
6	UBC 823	(TC)₈C	5	0	Pst-AG/Mse-AG	GACTGCGTACATGCAGAG GATGAGTCCTGAGTAAAG	54	0
7	UBC 825^*	(AC)₈T	7	28.6	Pst-AG/Mse-CG	GACTGCGTACATGCAGAG GATGAGTCCTGAGTAACG	36	0
8	UBC 827^*	(AC)₈G	12	8.3	Pst-CG/Mse-AG^*	GACTGCGTACATGCAGCG GATGAGTCCTGAGTAAAG	42	4.8
9	UBC 828	(TG)₈A	2	0	Pst-TC/Mse-TC	GACTGCGTACATGCAGTC GATGAGTCCTGAGTAATC	13	0
10	UBC 830	(TG)₈G	1	0	Pst-TC/Mse-AT	GACTGCGTACATGCAGTC GATGAGTCCTGAGTAAAT	20	0
11	UBC 834	(AG)₈CT	3	0	Pst-AT/Mse-AT	GACTGCGTACATGCAGAT GATGAGTCCTGAGTAAAT	50	0
12	UBC 840	(GA)₈CT	2	0	Pst-AT/Mse-TC	GACTGCGTACATGCAGAT GATGAGTCCTGAGTAATC	25	0
13	UBC 843	(CT)₈GA	1	0	Pst-AA/Mse-AA	GACTGCGTACATGCAGAA GATGAGTCCTGAGTAAAA	13	0
14	UBC 846	(CA)₈GT	3	0	Pst-AA/Mse-AC	GACTGCGTACATGCAGAA GATGAGTCCTGAGTAAAC	37	0
15	UBC 853	(TC)₈AT	6	0	Pst-AC/Mse-AC	GACTGCGTACATGCAGAC GATGAGTCCTGAGTAAAC	12	0
16	UBC 848	(CA)₈GG	-	-	Pst-AC/Mse-AA	GACTGCGTACATGCAGAC GATGAGTCCTGAGTAAAA	8	0
17	UBC 849	(GT)₈CA	-	-	Pst-GG/Mse-GG	GACTGCGTACATGCAGGG GATGAGTCCTGAGTAAGG	-	-
18	UBC 855	(AC)₈CT	4	0	Pst-GG/Mse-GA	GACTGCGTACATGCAGGG GATGAGTCCTGAGTAAGA	-	-
19	UBC 858	(TG)₈AT	-	-	Pst-GA/Mse-GA	GACTGCGTACATGCAGGA GATGAGTCCTGAGTAAGA	19	0
20	UBC 865^*	(CCG)₆	4	25.0	Pst-GA/Mse-GC	GACTGCGTACATGCAGGA GATGAGTCCTGAGTAAGC	9	0
21	UBC 867	(GGC)₆	-	-	Pst-CC/Mse-CC^*	GACTGCGTACATGCAGCC GATGAGTCCTGAGTAACC	20	30.0
22	UBC 875	(CTAG)₄	-	-	Pst-CC/Mse-GG	GACTGCGTACATGCAGCC GATGAGTCCTGAGTAAGG	43	0
23	UBC 881	(GGGT)₃G	-	-	Pst-TT/Mse-CC	GACTGCGTACATGCAGTT GATGAGTCCTGAGTAACC	48	0

* primers selected for analysis of genetic stability of micropropagated plants;

“-” no visible and scorable bands

Table 2

The post-effect of auxin type (IAA and IBA) and concentration (0.0; 1.0; 2.5; 4.0 mg·dm⁻³) on ex vitro acclimatization of Lonicera caerulea ‘Zojka’ and ‘Wojtek’ in the growth chamber

Genotype	Auxin type/concentration (mg·dm⁻³)		Survived micro-cutting (%)	Fresh mass of shoots (mg)	Shoot length (mm)	Fresh root mass (mg)	Root number/cutting	Root length (mm)
‘Zojka’	IBA	0.0	84 a	184 ± 46.9bc	115 ± 11.0bc	52.6 ± 20.7c	2.5 ± 0.9c	38.5 ± 9.8ab
		1.0	88 a	247 ± 69.7a	154 ± 21.7a	72.2 ± 35.0b	3.7 ± 1.0ab	37.1 ± 10.7ab
		2.5	80 a	174 ± 68.8bc	126 ± 11.3b	63.6 ± 23.4bc	3.8 ± 0.7a	32.4 ± 15.8bc
		4.0	76 a	179 ± 63.5bc	116 ± 17.4bc	54.8 ± 11.9bc	3.7 ± 1.0ab	28.7 ± 7.3c

	IAA	1.0	80 a	219 ± 77.4ab	111 ± 16.3c	89.0 ± 36.7a	3.5 ± 1.1ab	42.0 ± 14.0a
		2.5	76 a	160 ± 67.9c	117 ± 15.0bc	48.4 ± 14.9 c	3.3 ± 0.9ab	36.2 ± 11.4a-c
		4.0	72 a	142 ± 67.1c	114 ± 22.6bc	52.6 ± 28.7c	3.1 ± 0.9bc	33.1 ± 7.1bc

‘Wojtek’	IBA	0.0	80 a	218 ± 85.1b	126 ± 32.4b	42.7 ± 20.4 d	2.5 ± 0.7d	51.3 ± 8.5b
		1.0	88 a	349 ± 95.4a	181 ± 23.1a	120.1 ± 43.5a	3.7 ± 1.1ab	44.3 ± 7.8bc
		2.5	84 a	198 ± 73.8b	135 ± 29.7b	75.6 ± 22.4bc	3.6 ± 0.9ab	37.1 ± 8.8cd
		4.0	68 a	160 ± 81.5b	124 ± 38.5b	74.8 ± 25.7bc	3.2 ± 0.7 b-d	33.3 ± 8.5d

	IAA	1.0	84 a	339 ± 50.3a	146 ± 32.4b	86.3 ± 34.7b	4.1 ± 1.2a	59.0 ± 18.9a
		2.5	76 a	210 ± 61.1b	145 ± 17.2b	70.8 ± 26.4bc	3.4 ± 1.1bc	30.5 ± 6.8d
		4.0	76 a	216 ± 58.4b	140 ± 39.0b	58.5 ± 21.8cd	2.9 ± 1.0cd	42.4 ± 14.7c

[i] Means in the columns followed by the same letters do not differ significantly according to Duncan's multiple range test at p = 0.05; the assessment of significance of differences was done for each genotype separately; means ± SD

Plantlets of blue honeysuckle growing in the greenhouse: A – *ex vitro* rooted shoots of ‘Wojtek’ after 6 weeks of growth on the peat; B – *ex vitro* rooted shoots of ‘Wojtek’ after 6 weeks of growth on the peat and perlite; C – plants of ‘Wojtek’ and ‘Zojka’ after 2 years of growth *ex vitro*

Table 3

The effect of microcutting type and soil substrate type on ex vitro rooting and acclimatization of Lonicera caerulea ‘Zojka’ and ‘Wojtek’ in the greenhouse

Cutting type	Soil substrate	Rooted/survived plants (%)	Fresh mass of shoots (mg)	Shoot length (mm)	Fresh root mass (mg)	Root number/cutting	Root length (mm)
‘Zojka’

Individual shoots	peat	88 a	91.9 ± 20.0c	81.4 ± 7.0a	22.0 ± 8.7 c	2.8 ± 1.1a	25.6 ± 5.6a
	peat + sand	80 a	52.9 ± 9.0d	62.3 ± 8.0c	11.0 ± 3.6d	2.7 ± 0.7a	24.4 ± 5.8a
	peat + perlite	84 a	62.0 ± 17.0cd	69.0 ± 14.0bc	15.4 ± 8.3cd	2.5 ± 0.7a	23.7 ± 7.1a

Multiplied clumps	peat	68 a	127.7 ± 41.0b	68.4 ± 14.0bc	22.8 ± 8.7 c	1.8 ± 0.5b	12.0 ± 1.9b
	peat + sand	72 a	139.3 ± 75.0ab	50.3 ± 12.0d	33.0 ± 21.3b	2.9 ± 1.5a	15.3 ± 6.5b
	peat + perlite	84 a	164.2 ± 88.0a	73.6 ± 22.9ab	49.6 ± 24.4a	3.0 ± 0.7a	21.8 ± 7.5a

‘Wojtek’

Individual shoots	peat	96 a	97.8 ± 20.0bc	106.8 ± 14.9a	20.9 ± 6.6c	2.4 ± 0.7b	27.1 ± 7.4a
	peat + sand	80 ab	72.4 ± 20.0c	85.8 ± 15.0b	12.9 ± 6.9c	1.9 ± 0.4c	29.4 ± 8.1a
	peat + perlite	88 a	114.2 ± 54.0bc	112.3 ± 29.0a	24.0 ± 9.5bc	1.9 ± 0.5c	21.8 ± 4.5b

Multiplied clumps	peat	60 b	133.9 ± 34.0b	66.6 ± 14.0cd	36.3 ± 19.6b	2.0 ± 0.5bc	22.0 ± 6.8b
	peat + sand	76 ab	103.4 ± 18.0bc	60.4 ± 11.0d	20.0 ± 7.1c	1.6 ± 0.7c	10.8 ± 2.9c
	peat + perlite	88 a	191.2 ± 43.0a	73.8 ± 30.0bc	71.8 ± 33.0a	3.1 ± 1.0a	28.4 ± 12.1a

[i] Note: See Table 2

Table 4

The genetic evaluation of acclimatized plantlets of Lonicera caerulea ‘Wojtek’ and ‘Zojka’ compared to mother plants

No.	Primer	Total number of bands	Number of polymorphic bands	Range of amplicon (pb)	Total number of bands	Number of polymorphic bands	Range of amplicon (pb)

		‘Zojka’			‘Wojtek’
1	UBC 825	9	1	500–1000	11	0	400–1000
2	UBC 810	6	0	600–1300	5	0	500–1300
3	UBC 827	12	0	300–800	11	0	300–750
4	UBC 865	9	0	300–1400	13	0	300–1400

Total		36	1		40	0

Proportion of polymorphic bands (%)		2,77			0,00

1	Pst-CC/Mse-CC	19	1	80–300	16	1	50–300
2	Pst-TT/Mse-CC	35	1	250–900	30	0	250–900
3	Pst-GC/Mse-TA	18	0	300–850	16	0	400–850
4	Pst-CG/Mse-AG	42	0	200–760	40	0	200–750

Total		105	2		102	1

Proportion of polymorphic bands (%)		1,90			0,98

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DOI: https://doi.org/10.2478/johr-2020-0019 | Journal eISSN: 2353-3978 (formerly 2300-5009) | Journal ISSN: 2300-5009

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Language: English

Page range: 61 - 70

Submitted on: May 1, 2020

Accepted on: Sep 1, 2020

Published on: Dec 31, 2020

Published by: National Institute of Horticultural Research

In partnership with: Paradigm Publishing Services

Keywords:

AFLP,

ISSR,

Related subjects:

Ecology,

© 2020 Agnieszka Wojtania, Monika Markiewicz, Justyna Góraj-Koniarska, published by National Institute of Horticultural Research
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Volume 28 (2020): Issue 2 (December 2020)

Ex vitro Rooting, Acclimatization and Genetic Stability of Lonicera caerulea var. kamtschatica

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