Aspectos citológicos das interações compatíveis e incompatíveis entre mudas de cajueiro (<i>Anacardium occidentale</i> L.) e isolados do complexo <i>Colletotrichum gloeosporioides</i> (Penz.)

Lopez, Ana Maria Queijeiro; Lucas, John Alexander

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Revista Brasileira de Fruticultura

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Plant Protection • Rev. Bras. Frutic. 43 (2) • 2021 • https://doi.org/10.1590/0100-29452021701 copiar

Aspectos citológicos das interações compatíveis e incompatíveis entre mudas de cajueiro (Anacardium occidentale L.) e isolados do complexo Colletotrichum gloeosporioides (Penz.)

Autoria SCIMAGO INSTITUTIONS RANKINGS

Resumo

Uma estratégia no controle da antracnose do cajueiro (Anacardium occidentale L.) é o manejo da fenologia e mecanismos de defesa deste hospedeiro. Em estudos anteriores, em condições controladas, as reações de mudas de 5 clones de caju (CAP-14, CCP-06, CCP-09, CCP-76 e CCP-1001) a 36 isolados do complexo Colletorichum gloeosporioides Penz. (LARS-905 a 940), foram avaliadas. Contudo, um bom manejo em campo requer informações sobre o processo de infecção. Esta pesquisa visou a avaliar aspectos citofisiológicos de três interações compatíveis deste patossistema (isolados LARS-905 e 910 × CCP-76; LARS-910 × CCP-1001) e uma incompatível (LARS-905 × CCP-1001), utilizando folhas/caules infectados e técnicas de microscopia (luz, varredura e transmissão eletrônica). Nenhuma diferença significativa foi encontrada antes da penetração. Nas combinações suscetíveis, 36-66 h após a inoculação, uma delgada hifa primária (TPH) formou-se na célula epidérmica invadida, alargando-se como uma larga hifa primária (LPH), que preencheu o lúmen da célula, simultaneamente com o acúmulo de lignopolissacarídeos amarelo-acastanhados. Então, uma fina hifa secundária (TSH) desenvolveu-se a partir do LPH, penetrando nas células adjacentes antes que as primeiras se tornassem necróticas. Na interação incompatível, a resposta da primeira célula invadida foi mais rápida e intensa, com papila e complexo lignopolissacárico-proteico-siliconoso geralmente bloqueando o patógeno.

Termos para indexação
antracnose; mecanismos de defesa; fungo hemibiotrófico; papila; resistência

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[TFN0101] ^a
A minimum of 100 conidia, appressoria or reaction sites were recorded in each of 3 replicates of 5 samples per interaction per time. b Percentage from the germinated conidia. c Percentage from the appressoria formed and/or attempts of penetration. Papillae (mainly dome-like shape) were observed during cuticular penetration and thin primary hypha (TPH) formation, in all the interactions. d Percentage from attempts of penetration (with or without papillae). LPH formed in the absence of papilla were more common in compatible interactions. e Percentage from the LPH formed; f When multiple penetration attempts occurred in the same cell, its rapid collapse restricted the TPH advance; g Penetration restricted to the first cell containing LPH; h Because of the rapid and intense browning of the invaded cells, SH were not recorded in whole tissues, but in semi-thin resin embedded sections (LM). Gl. Slide = glass slide; conidiat. = conidiation.

Inoculation	Development Phase	910 × CCP76	910 × CCP1001	910 × CAP14	905 × CCP76	905 × CCP1001	905 × CAP14	910	905
12	Germination Appressorium formed^b	93 75	92 77	90 72	91 73	89 72	94 76	90 72	91 71
18	Germination Appressorium formed^b Papillae^c	95 86 21	94 85 20	96 83 23	97 82 23	90 80 27	95 83 24	95 87 -	93 85 -
24	Appressorium formed^b Papillae^c	90 22	89 23	90 26	89 25	88 30	93 27	91 -	89 -
38	Papillae^c	36	38	40	39	42	40	-	-
48	Papillae^c Large Primary ***** (LPH)^d	47 16	47 17	48 18	49 17	52 13	52 19	- -	- -
66	Papillae^c LPH^d	69 33^f	71 32^f	71 34^f	73 32^f	75 17^g	73 35^f	- -	- -
74	LPH^d ******** of other cells Secondary hypha (SH)^e	37^f 2-3 cells 15	38^f 2-3 cells 14	37^f 2-3 cells 14	35^f 2-3 cells 13	23^g 1-2 cells^g ^g	31^f 2-3 cells 15	- - -	- - -
96	Invasion of ***** cells SH^e	3-5 cells 18^h	3-5 cells 16^h	3-5 cells 17^h	3-5 cells 17^h	1-2 cells^g ^g	3-5 cells 19^h	-	-
114	lesion development Growth in necrotic tissues	+ +	+ conidiat. (acervuli and setae)	+ conidiat. (******** and setae)	+ +	- -	+ +	- -	- -

Inoculation	Development Phase	910 × CCP76	910 × CCP1001	910 × CAP14	905 × CCP76	905 × CCP1001	905 × CAP14	910	905
12	Germination Appressorium formed^b	91 75	89 73	93 68	89 70	85 69	92 72	90 72	91 71
18	Germination Appressorium formed^b Papillae^c	94 81 23	92 83 25	96 80 21	93 80 25	91 79 29	96 82 23	95 87 -	93 85 -
24	Appressorium formed^b Papillae^c	88 29	86 33	92 30	90 31	89 36	94 32	91 -	89 -
38	Papillae^c	39	41	42	41	47	44	-	-
48	Papillae^c Large Primary Hypha (LPH)^d	50 20	52 22	53 25	52 21	57 17	53 23	- -	- -
66	Papillae^c LPH^d	71 37^f	72 35^f	73 36^f	73 37^f	78 21^f	75 35^f	- -	- -
74	LPH^d Invasion of other cells ********* hypha (SH)^e	43^f 2-3 cells 20	40^f 2-3 cells 19	42^f 2-3 cells 23	41^f 2-3 cells 18	25^g 1-3 cells^g ^g	41^f 2-3 cells 20	- - -	- - -
96	Invasion of other cells SH^e	3-5 cells 23^h	3-5 cells 22^h	3-5 cells 25^h	3-5 cells 22^h	1-3 cells^g ^g	3-5 cells 21^h	-	-
114	lesion development Growth in necrotic tissues	+ +	+ conidiat. (******** and setae)	+ conidiat. (acervuli and setae)	+ +	- -	+ conidiat. (acervuli and setae)	- -	- -

		Reaction
Aniline blue (AB)	Callose = green-yellow fluorescence	+
AB- ********* blue (TB)	Callose = green-yellow fluorescence	+
Autofluorescence	Lignin = fluorescence	-
TB	Lignin = blue-green Callose = unstained Hemicellulose, cellulose, pectin = purple-red Silicates and polysulfates = purple-red Proteinaceous material = purple-red	Purple-red
TB - CaCl₂	Lignin = blue; other ********* = unstained	Unstained*
Phloroglucinol - HCl	Guayacyl lignin = red Syringaldeyde lignin = unstained	Unstained*
Phloroglucinol:CaCl₂-HCl	Lignin = deep orange ** red	Unstained*
Chlorine water: Na₂SO₃	Guayacyl lignin = brown Syringyl lignin = red Catechol, resorcinol, quinol = unstained Tannic acid, methyl gallate, pyrogallol = pink	Unstained*
KMnO₄ - *** - NH₄OH	Siringyl-******* lignin = purple to red-brown	Unstained*
Diazotized p-nitroaniline	Phenols with an ortho para free position = yellow-orange to brown *** and sinapic acids = pink Lignin = orange-* to red	Unstained*
p-nitroaniline-tetrafluorborate	Phenols with an orthoor para free position = orange-red.Red-violet after NaOH	Unstained*
FeCl₃	Pectin-tannin, hydroxamic acid derivatives = reddish-brown; Catechol = blue-green	reddish-brown (healthy cells = blue)
Nitroso Reaction	Anthocyanins, anthoxanthins = yellow Gallotannins= yellow,orange-red Flavonols, ellagitannins, catechins and catechol derivatives (hydroxynaftoquinones)= cherry to red-brown	Deep Purple
Vanillin: HCl	Condensed tannins, dihydrochalcones, anthocyanins, resorcinol = pink to brick-red Catechol, pyrogallol, hydroquinone, phloroglucinol, hydrolisable tannins = unstained	Brick red (fading to orange). Red in blue ***** excitation (filter barrier 520 nm)
Syringaldazine- H₂O₂	Laccase = ****** before adding H₂O₂ Peroxidase = purple after adding H₂O₂	Purple with H₂O₂
Coomassie Blue (Cb)	Acid and neutral proteins = blue violet Histones, PRPs, HPRPs = purple-red	Purple-red
NaOH (0.5-10 N)	Dissolves protoplasm; saponifies suberin Cyanidin *********** = blue	Unstained* (***** *** = blue)
H₂O₂: CH₃COOH	Dissolves brown quinones	Pale-yellow

	Penetration attempts(appressoria formed)	******** at penetration attempts	*********** without papillae	Penetration with Papillae	papillae or penetration
Cycloheximide	1432 1398	1102 58	141 1129	98 84	92 127