<i>In vitro</i> pollination and fluorescence microscopy for characterization of gametophytic self-incompatibility of C<i>offea canephora</i> Pierre ex A. Froehner

Souza, Carolina Augusto de; Rocha, Rodrigo Barros; Santos, Maurício Reginaldo Alves dos; Lopes, Tatiane Almeida; Teixeira, Alexsandro Lara; Espindula, Marcelo Curitiba

doi:10.1590/1984-70332021v21n4a51

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Crop Breeding and Applied Biotechnology

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ARTICLE • Crop Breed. Appl. Biotechnol. 21 (4) • 2021 • https://doi.org/10.1590/1984-70332021v21n4a51 copy

In vitro pollination and fluorescence microscopy for characterization of gametophytic self-incompatibility of Coffea canephora Pierre ex A. Froehner

Authorship SCIMAGO INSTITUTIONS RANKINGS

Abstract

The objectives of this study were to optimize in vitro pollination and fluorescence microscopy procedures to characterize the gametophytic self-incompatibility of Coffea canephora genotypes. For that purpose, 115 in vitro pollinations were carried out among 30 parent plants of the Conilon (17) and Robusta (13) botanical varieties. Tester plants of compatibility groups I, II, and III were used as pollen grain donors. On the day of anthesis, pollen grains of the donor genotypes were transferred in vitro to one of the sides of the bifid stigma of each receptor flower. After 30 hours, the development of the pollen tubes was evaluated in fluorescence microscopy. Eleven genotypes (37%) were classified in compatibility Group I, thirteen (43%) in Group II, and six (20%) in Group III. In vitro pollination allowed reduction in the time necessary to diagnose compatibility, without exhibiting contamination caused by the presence of pollen grains of non-donor plants.

Keywords:
Canephora coffee; Conilon; Robusta; compatibility

n	Clone	Tester hybridizations			FN	FP	Group	p value
n	Clone	I	II	III	FN	FP	Group	p value
1	C 125	2	(1)	2	0	0	II	0.05
2	C 154	(2)	1	1	0	0	I	0.05
3	C 167	(1)	1	1	0	0	I	0.05
4	C 184	1	(1)	1	0	0	II	0.05
5	C 452	(2)	1	1	0	0	I	0.01
6	C 46	1	(1)	2	0	0	II	0.05
7	C468	1	(1)	2	0	0	II	0.05
8	C 484	1	1	(1)	0	0	III	0.05
9	C 59	1	(1)	1	0	0	II	0.05
10	C 694	2	1	(1)	0	0	III	0.05
11	C 729	1	(1)	2	1	0	II	0.05
12	C 795	1	(1)	1	1	0	II	0.05
13	C 799	(2)	1	1	0	0	I	0.01
14	C 801	1	(1)	1	0	0	II	0.05
15	C 945	1	(1)	1	0	0	II	0.05
16	C 968	1	1	(1)	0	0	III	0.05
17	C 986	(1)	1	1	0	0	I	0.05
18	R 164	2	2	(1)	0	0	III	0.01
19	R 183	(1)	2	1	1	0	I	0.05
20	R 214	(1)	1	1	0	0	I	0.05
21	R 216	(1)	1	1	1	0	I	0.05
22	R 277	1	(1)	1	0	0	II	0.05
23	R 3	(1)	1	2	0	0	I	0.05
24	R 30	(1)	4	1	0	0	I	0.05
25	R 32	3	1	(2)	0	0	III	0.05
26	R 36	2	1	(3)	0	0	III	0.05
27	R 5	(1)	1	1	0	0	I	0.05
28	R 63	1	(1)	1	0	0	II	0.05
29	R 9	1	(1)	1	0	0	II	0.05
30	R 99	3	(1)	2	0	0	II	0.01

In vitro pollination	I	II	III	Total
Compatible hybridizations	27	22	30	79
Non-compatible hybridizations	14	13	9	36
Total hybridizations	41	35	39	115
False negative	4
False positive	0
False negative rate	3.45

Type of error	Source of error	Form of control
False negative	Viability of the pollen grains	Performance of germination test prior to in vitro pollination
	Transfer of the pollen grains	Training for development of manual ability in scraping pollen grains
	Stigma maturity	Collection of stigmas in the field and transfer to the culture medium on the day before anthesis
False positive	Contamination of pollen grains	Use of asepsis procedures during in vitro pollination
	Mistaken diagnosis	Training for visual recognition of the pollen tubes Careful preparation of the slides
	Error in identification of the samples	Identification of the samples in all the steps of evaluation and control of the process

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