Co esponding au ho : Bi T a Jean-Innocen NANTI
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In i o egene a ion o cashew [Anaca dium occiden ale L. (Anaca diaceae)] ees
om g a ed plan s
Bi T a Jean-Innocen NANTI 1, *, Yah Gwladis GNAMIEN 1, A oué Rachelle N’GUESSAN 1, Konan Ma ius
KOUASSI 2 and Mongomaké KONÉ 2
1 Labo a o y o he Imp o emen o Ag icul u al P oduc ion, UFR Ag o o es y, Jean Lo ougnon Guédé Uni e si y, BP 150
Daloa, Cô e d’I oi e.
2 Labo a o y o Biology and Imp o emen o Plan P oduc ion, UFR Na u al Sciences, Nangui Ab ogoua Uni e si y, 02 BP
801 Abidjan 02, Cô e d’I oi e.
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 27(01), 1593-1601
Publica ion his o y: Recei ed on 13 May 2025; e ised on 15 July 2025; accep ed on 17 July 2025
A icle DOI: h ps://doi.o g/10.30574/wja .2025.27.1.2563
Abs ac
P opaga ion by seeds o he cashew ee (Anaca dium occiden ale L.) does no allow a ai h ul ep oduc ion o he
mo he plan . The objec i e o his wo k is o es ablish an in i o cashew ee egene a ion p o ocol om he apex o
g a ed plan s. To his end, apex explan s aken om plan s g a ed in g eenhouses we e disin ec ed using sodium
hypochlo i e wi h 1.2 and 2.4% ac i e chlo ine and calcium hypochlo i e 2.5 and 5% o 1; 2 ; 3 and 5 min, hen cul u ed
on MS medium supplemen ed wi h benzylaminopu ine (BAP), kine in and hidiazu on (TDZ) a a ious concen a ions.
The main esul s showed ha disin ec ion o he explan s wi h sodium hypochlo i e a ac i e chlo ine 1.2 % o 2 min
esul ed in a low con amina ion a e (33%). The budding a e and he mean numbe o buds (1 o 2 buds pe explan )
did no a y while he bes shoo g ow h (0.4 cm) we e ob ained on he medium wi hou g ow h egula o s. A he end
o hese expe imen s, he p o ocol o in i o egene a ion om he apex o he cashew ee was es ablished.
Keywo ds: Anaca dium occiden ale; In i o; Disin ec ion; Regene a ion
1. In oduc ion
Na i e o La in Ame ica, he cashew ee (Anaca dium occiden ale L.) was in oduced o Cô e d'I oi e in 1951. I was
only om 1959 o 1960 ha cashew ee o es plan a ion p og ammes we e ca ied ou and ex ended o he en i e
ecologically a ou able a ea, in pa icula he Sudano-Guinean sa annahs [1]. The objec i e a he ime was he
p o ec ion o ecosys ems se iously a ec ed by de o es a ion and he igh agains bush i es [2]. In 1990, he inc ease
in he p ice o cashew nu s led o a c aze o he cul i a ion o cashew ees (Anaca dium occiden ale L.) [3]. As a esul ,
cashew ees appea ed o p oduce s as an al e na i e o he adi ional cash c op o he no h o he coun y, co on,
whose cul i a ion seemed h ea ened by alling global cos s and majo p oblems in he unc ioning o he sec o [4].
The cashew ee is cul i a ed o he ade o i s nu s. I is used as a aw ma e ial in he ag i- ood (chocola e,
con ec ione y), pha maceu ical and cosme ic indus ies [5]. Cashew apples a e also app ecia ed because o hei
ichness in wa e (80 o 90%) and suga s (10 o 12%). This alo iza ion o apple juice has been he subjec o se e al
s udies [6,7].
I is es ima ed ha be ween 1980 and 2020, he o al a ea o land de o ed o cashew cul i a ion inc eased om 526,250
hec a es (ha) o 7,101,970 ha. As o wo ld p oduc ion, om 706,500 ons in 1990, i inc eased om 3.9 million ons in
2018 o mo e han 5,000,000 ons in 2022. This expansion has been obse ed mainly in A ica. Cô e d'I oi e has been
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 27(01), 1593-1601
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he epicen e o his boom in global cashew p oduc ion. This Wes A ican coun y is now he wo ld's la ges expo e
o aw cashew nu s, wi h a u no e o $961 million in 2021 and a p oduc ion o mo e han one million ons in 2022 [8].
This inc ease in na ional p oduc ion is due o he inc ease in cul i a ed a eas, which ose om 265,654 ha in 2000 o
1,350,000 ha in 2018 [9]. This galloping inc ease in cul i a ed land can lead o a lack o insu iciency o a able land and
land dispu es. Nu yields in I o ian o cha ds emain low, in he ange o 350 o 500 kg/ha, due o plan a ions c ea ed
wi h unimp o ed plan ma e ial [10]. Con en ional ege a i e p opaga ion echniques (laye ing, cu ings, g a ing)
ha e been used o he p oduc ion o high-p oducing cashew plan ing ma e ial [11]. These me hods allow he mo he
plan o be ai h ully ep oduced, bu emain slow p opaga ion ools [12]. In addi ion o hese echniques, in i o cul u e
me hods o plan issues and o gans ha e been used [13; 14]. Bio echnology is a powe ul ool o o e coming na u al
ba ie s, and hus ep esen s an al e na i e and complemen a y app oach o con en ional b eeding e o s [15].
Un o una ely, he mic op opaga ion o woody species om explan s aken om ma u e ees is s ongly hampe ed by
se ious cons ain s such as he di icul y o disin ec ion and b owning o he explan s and he low a e o bud induc ion,
hus making hei mul iplica ion di icul in i o. Despi e hese cons ain s, au ho s ha e epo ed he p oduc ion o
cashew nu plan ma e ial h ough mic op opaga ion [16; 13]. In addi ion, he condi ions o g owing hese explan s
a e no clea ly es ablished. The objec i e o his s udy is o es ablish a egene a ion p o ocol o he p oduc ion o
cashew nu i oplan s om g a ed plan s.
2. Ma e ials and me hods
2.1. Plan ma e ial
The plan ma e ial consis s o young cashew plan s h ee mon hs old ob ained by g a ing (Figu e 29). The g a s we e
aken om high-yielding ees in an indus ial plan a ion in Fa ako, Bogo a Sub-P e ec u e, Gon ougou egion (Cô e
d'I oi e). Roo s ocks a e ob ained by ge mina ing ma u e nu s ha es ed in Gohi a la, in he Ma ahoué egion (Cô e
d'I oi e).
Figu e 1 Th ee-mon h-old g a ed cashew ee plan
2.2. P epa a ion o cul u e media
The base medium o Mu ashige and Skoog [17] was used o his s udy. To his medium, 30 g/l suc ose and an
an ioxidan (ac i a ed cha coal 0.2%) and a ious concen a ions o cy okinins we e added. The pH was adjus ed o 5.8
using a pH me e wi h HCl (1N) and/o NaOH (1N). The gelling agen , phy agel (3g/l) has been added. The esul ing
solu ion was s i ed o 10 min and hen b ough o a boil in o de o solubilize he phy agel. A olume o 30 ml o his
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 27(01), 1593-1601
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medium was dispensed in o po s which ha e been he me ically sealed wi h caps. These media we e s e ilized a
au ocla e a 121 °C o 20 min unde a p essu e o 1 ba and hen ans e ed o a oom o cul u e a e cooling.
2.3. P epa a ion and disin ec ion o explan s
G a ed plan s ea ed wi h ungicide once a week we e used as a sou ce o explan s. The wigs aken ea ly in he
mo ning om hese plan s we e soaked in soapy wa e o 30 minu es and hen insed ho oughly wi h ap wa e . Unde
he lamina low hood, hese wigs a e imme sed in 70% alcohol o 1 min, hen in a ious concen a ions o sodium o
calcium hypochlo i e con aining 2 d ops o ween 20 a di e en imes.
2.4. Cul i a ion o explan s
A e he disin ec ion phase, he apex explan s we e aken om he disin ec ed wigs and placed on he MS medium
supplemen ed wi h 0.2% ac i a ed cha coal and a ious concen a ions o cy okinins
Figu e 2 Cul u ing he apex
2.5. E ec o disin ec ing agen s on explan con amina ion and iabili y
Sodium hypochlo i e a 1.2 and 2.4% ac i e chlo ine and calcium hypochlo i e 2.5 and 5% we e he disin ec ing agen s
used. In hese solu ions, he explan s we e soaked a 1; 2 ; 3 and 5 min. A he end o he imme sion ime, he explan s
we e emo ed and insed h ee imes wi h s e ile dis illed wa e . The iabili y o he explan s was e alua ed a e h ee
weeks o cul i a ion. The expe imen was epea ed h ee imes a a a e o 10 eplica ions pe ea men and pe epea .
2.6. E ec o cy okinins on cashew bud induc ion
The apex explan s we e cul u ed on MS medium supplemen ed wi h h ee cy okinins a di e en concen a ions. Thus,
benzylaminopu ine (BAP) and kine in a concen a ions o 0, 1, 2, 4 and 6 mg/L and hidiazu on (TDZ) a concen a ions
o 0; 0.001, 0.01, 0.1 and 1 mg/L we e used as g ow h egula o s. The apex explan s we e placed on hese di e en
media. The expe imen was epea ed h ee imes o each medium, a a a e o 10 eplica ions pe epea
2.7. Expe imen al design and da a analysis
The expe imen s we e ca ied ou in a comple ely andomized design wi h en eplica es and each indi idual ea men
was epea ed h ee imes. Bud equency, numbe o buds pe explan we e submi ed o analysis o a iance (ANOVA)
o de ec signi ican di e ences be ween means o each g ow h egula o and disin ec an . Means di e ing signi ican ly
we e compa ed using Newman-Keuls mul iple ange es a .
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3. Resul s
3.1. E ec o disin ec an s and hei concen a ions and du a ion o disin ec ion on explan con amina ion
The esul s ega ding he e ec o disin ec an s a e p esen ed in able I. Analysis o he esul s in his able showed a
e y signi ican e ec o disin ec ing agen s on he con amina ion a e (P < 0.001). This is because his con amina ion
a e is highe wi h calcium hypochlo i e compa ed o sodium hypochlo i e, ega dless o he du a ion o disin ec ion. In
addi ion, his a e dec eases wi h inc easing disin ec ion ime. Thus, he highes con amina ion a es a e 87.09% and
90% espec i ely a 1 min and 2 min wi h sodium hypochlo i e (1.2% ac i e chlo ine) and calcium hypochlo i e (2.5%).
Table 1 Con amina ion a e o cashew ee explan s a e disin ec ion wi h sodium and calcium hypochlo i e
Disin ec an s
Concen a ions (%)
Du a ion (min)
Con amina ion a e
Sodium
hypochlo i e
1.2
1
87.09 ± 2,9 a
2
33 ± 8.05 cd
3
30 ± 7.87 cd
5
18 ± 6.68 d
2.4
1
78 ± 5.68 a
2
18 ± 6.68 d
3
24 ± 7.39 d
5
12 ± 5.68 d
Calcium
hypochlo i e
2.5
1
90 ± 00 a
2
90 ± 00 a
3
72 ± 6.68 ab
5
63 ± 7.65 ab
5
1
72 ± 6.68 ab
2
51 ± 8.28 ab
3
66 ± 7.39 ab
5
33 ± 8.05 cd
P obabili y
P1
P2
P3
P4
< 0.001
< 0.001
< 0.001
0.47
In he same column, he means ollowed by he same le e a e signi ican ly iden ical o he
5% h eshold (S uden Newman-Keuls es ). Mean ± s anda d e o (P1): p obabili y ela ed
o he na u e o he disin ec an agen ; (P2): p obabili y ela ed o he concen a ions o
disin ec an s; (P3): p obabili y ela ed o he du a ion o disin ec ion; (P4): p obabili y o
he disin ec an agen -concen a ion-du a ion o disin ec ion in e ac ion.
3.2. E ec o disin ec ing agen s on he iabili y o he explan
A e one mon h o cul u e o he apexes, he a e o explan s ha su i ed he disin ec an s is shown in able 2. Analysis
o his able e eals ha he disin ec ing agen did no ha e a signi ican e ec on explan iabili y (P > 0.05). In con as ,
concen a ions and du a ion o disin ec ion had a e y signi ican e ec on explan su i al (P < 0.001). These
concen a ions anged om 18% ob ained wi h sodium hypochlo i e a 2.4% ac i e chlo ine du ing 5 min o
disin ec ion, o 87% ob ained wi h sodium hypochlo i e a 1.2% ac i e chlo ine du ing 1 min o disin ec ion. Taking in o
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 27(01), 1593-1601
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accoun he con amina ion a e and su i al a e, sodium hypochlo i e wi h 1.2% ac i e chlo ine used o 2 min was he
mos indica ed o e ec i e disin ec ion o he apex explan s o he cashew ee.
Table 2 Cashew apex su i al a e a e disin ec ion wi h sodium calcium hypochlo i e
Disin ec an s
Concen a ions (%)
Du a ion (min)
Viabili y a e
Sodium hypochlo i e
1.2
1
87.09 ± 2.9 a
2
81 ± 5.01 a
3
48 ± 8.34 bcd
5
33 ± 8.05 cd
2.4
1
66 ± 7.39 ab
2
51 ± 8.28 bc
3
30 ± 7.88 cd
5
18 ± 6.68 de
Calcium hypochlo i e
2.5
1
83.79 ± 4.31 a
2
66 ± 7.39 ab
3
48 ± 8.34 bc
5
48 ± 8.34 bc
5
1
72 ± 6.68 ab
2
45 ± 8.36 bcd
3
42 ± 8,34 bcd
5
6,33 ± 4,17 e
P obabili y
P1
P2
P3
P4
0,916
< 0.001
< 0.001
0,171
In he same column, he means ollowed by he same le e a e signi ican ly iden ical o he 5% h eshold (S uden
Newman-Keuls es ). Mean ± s anda d e o (P1): p obabili y ela ed o he na u e o he disin ec an agen ; (P2):
p obabili y ela ed o he concen a ions o disin ec an s; (P3): p obabili y ela ed o he du a ion o disin ec ion;
(P4): p obabili y o he disin ec an agen -concen a ion-du a ion o disin ec ion in e ac ion.
3.3. E ec o cy okinins on cashew budding
A e a mon h o cul i a ion, he cul i a ed apexes ha e budded. These buds u ned in o lea y shoo s a e six mon hs o
cul i a ion (Figu e 3). Values o he e ec o bensylaminopu ine, kine in and hidiazu on on budding ha e been
epo ed in Table 3. Analysis o he esul s shows ha he cy okinins used ha e a simila e ec on budding ime, budding
a e, numbe o buds and mean shoo size (P > 0.05). Howe e , cy okinin concen a ion had a signi ican in luence on
budding ime and mean bud coun (P < 0.05) and a e y signi ican e ec on mean bud size (P < 0.001). No cy okinin-
concen a ion in e ac ion e ec was obse ed on he a ious pa ame e s obse ed (P > 0.05). The i s bud appea s
eigh days a mos a e he cul i a ion o he explan s; The budding a e anged om 39% o 61%. In addi ion, he
a e age numbe o buds pe explan did no exceed 2. Rega ding he a e age size o he shoo s, i a ied om 0.28 cm
on medium supplemen ed wi h TDZ 1mg/L o 0.42 cm on medium lacking cy okinin.
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 27(01), 1593-1601
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Table 3 Pa ame e s o budding o cashew apex acco ding o cy okinins and hei concen a ions
Cy okinins
Concen a ions (mg/L)
Budding ime
(day)
Budding a e
(%)
buds
numbe
A e age
heigh
BAP
1
6.87 ± 0.17 a
42 ± 8.33 a
1.4 ± 0.09 a
0.42 ± 0.02 a
2
7.3 ± 0.19 a
54 ± 8.18 a
1.6 ± 0.1 a
0.40 ± 0.01 ab
4
7.3 ± 0.23 a
60 ± 7.87 a
1.6 0.09 a
0.37 ± 0.01 abc
6
7.43 ± 0.18 a
48 ± 8.33 a
1.5 ± 0.09 a
0.31 ± 0.01 cd
KIN
1
6.97 ±0.18 a
45 ± 8.35 a
1.57 ± 0.1 a
0.4 ± 0.02 ab
2
6.9 ±0.22 a
54 ± 8.1 a
1.5 ±0.09 a
0.37 ± 0.01 abc
4
7.37 ± 0.25 a
54 ± 8.18 a
1.43 ± 0.09 a
0.38 ± 0.01 abc
6
7.6 ± 0.22 a
54 ±8.18 a
1.4 ±0.09 a
0.31 ± 0.1 cd
TDZ
0.001
7.4 ± 0.20 a
60 ± 7.87 a
1.57 ± 0.12 a
0.39 ± 0.01 abc
0.01
7.33 ± 0.19 a
66 ± 7.39 a
1.77 ± 0.12 a
0.37 ± 0.01 abc
0.1
7.63 ± 0,17 a
54 ± 8.18 a
1.73 ± 0.11 a
0.35 ± 0.01 abc
1
7,67 ±0.18 a
57 ± 8.05 a
1.57 ± 0.10 a
0.28 ± 0.01 d
P obabili y
P1
0.059
0.597
0.050
0.334
P2
0.003
0.142
0.037
< 0.001
P3
0.828
0.905
0.848
0.965
In he same column, he means ollowed by he same le e a e signi ican ly iden ical o he 5% h eshold (S uden Newman-Keuls es ). Mean ±
s anda d e o (P1): cy okinin- ela ed p obabili y; (P2): p obabili y ela ed o cy okinin concen a ions; (P3): p obabili y o cy okinin-BAP
concen a ions: Benzylaminopu ine; Kin: Kine ine; TDZ: Thidiazu on
Figu e 3 Bud induc ion and lea y shoo de elopmen om he explan apex aken om g a ed plan s(a) bud
induc ion; (b) bud de elopmen in o lea y shoo s (b g): buds
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4. Discussion
Exogenous and endogenous con amina ion o explan s has always conside ably limi ed he in i o cul u e o plan s.
Mic oo ganisms, including bac e ia and ungi, as well as some mi es ha ca y hem, can be associa ed wi h explan s
used o in i o plan egene a ion [18]. Thus, one o he possible easons o in ec ion o explan s could be he o igin
o he plan ma e ial used. [19] sugges ed ha mo he plan s used o plan issue cul u es should be g own unde
con olled condi ions (in g eenhouses o g ow h chambe s) o minimize he isk o mic obial con amina ion o o gans.
Howe e , ega dless o he o igin o he explan s, he use o good disin ec ion echniques, depending on he plan o gan
o issue conside ed, should allow he elimina ion o mic obial con amina ion [20]. The losses obse ed a y depending
on he na u e o he disin ec ing agen , he concen a ion and he du a ion o disin ec ion. In his s udy, wo chemical
solu ions we e es ed o hei abili y o neu alize mic oo ganisms ha in ec cashew ee explan s
The la ges numbe o disin ec ed explan s, bo h o which su i ed he e ec o he chemical agen s, was ob ained a e
a 2-minu e soak in sodium hypochlo i e wi h 1.2% ac i e chlo ine. This high su i al a e wi h his disin ec ing agen
a es s ha sodium hypochlo i e disin ec ion is e ec i e o he su ace s e iliza ion o he apex explan s o he cashew
ee. The e ec i eness o sodium hypochlo i e makes i he mos widely used disin ec an [21; 22] o in i o ini ia ion
o explan s, compa ed o calcium hypochlo i e. [23] showed ha sodium hypochlo i e used a 3.6% ac i e chlo ine is
e ec i e in disin ec ing Lippia mul i lo a explan s. Indeed, sodium hypochlo i e seems o pene a e inside cells and
he e o e ac s mo e e ec i ely on endogenous pa hogens in cells. In addi ion, sodium hypochlo i e, due o i s dual
an i ungal and an ibac e ial ac ion, has been shown o signi ican ly educe ungal and bac e ial in ec ions [24].
A e explan s disin ec ion phase, he s udy o he in luence o cy okinins on budding showed ha he numbe o buds
pe explan a ied be ween 1 and 2. [25] showed he low po en ial o induc ion o he apex bud compa ed o nodal
segmen s. The p oduc ion o a bud by in i o cul u e om cashew explan s om an ex i o sou ce was epo ed by
[26], a e wo mon hs o cul u e. This au ho ob ained a single bud pe apex explan on he MS and LS media. The low
o ganogenic po en ial o he apex was epo ed by [27]. These esea che s ob ained a maximum o wo buds pe explan
(apex) when hey wo ked on he mic op opaga ion o cashew nu s. [28] did no obse e any buds on he apex o he
explan collec ed om plan s ob ained by ex i o ge mina ion. Howe e , o he au ho s ha e epo ed ob aining a
g ea e numbe o buds in in i o cul u e o he apex o he cashew nu . [29] obse ed wo o h ee axilla y shoo s pe
explan du ing cashew mic op opaga ion, while [30] ob ained h ee o ou buds pe explan . The low numbe o buds
pe explan ob ained in his s udy compa ed o some p e ious wo k may be due o he geno ype o he plan s used as a
sou ce o explan s. Indeed, mos plan s exhibi species-speci ic geno ypic egene a ion. Wi hin he same species, one
geno ype p oduces buds while ano he can only p oduce emb yos [31]. Howe e , se e al au ho s men ion ha only
ce ain geno ypes seem o ha e a high o ganogenic powe . In many species, his abili y appea s o be con olled by
geno ype [32]. Such con ol o egene a ion has been epo ed in legumes, pa icula ly in al al a by [33], in T i olium
epens L. by [34] and many o he species. [35] indica e he p esence o a ew majo genes ha can con ol o ganogenesis
in peas. Simila ly, [36] succeeded in signi ican ly inc easing he emb yogenic equency in Médicago sa i a L. a e wo
s ages o ecu en selec ion. [37] ound in a diploid al al a geno ype ha bud di e en ia ion om callus is con olled
by wo dominan genes called Rn1 and Rn2, whose simul aneous p esence allows o a high egene a ion a e (mo e
han 75% o explan s). The budding a e is abou 60%. This esul co obo a es hose o [13] who ob ained a budding
a e o 57.2% in in i o cashew cul u e om explan s aken om plan s g own in g eenhouses. Se e al o he au ho s
ha e ob ained 80-100% budding in he same species wi h high concen a ions o sodium hypochlo i e [38; 25]. The low
budding a es eco ded in his s udy could be explained by he ac ha chemical solu ions used a low concen a ions
ha e di icul y pene a ing he explan s. These a e unable o neu alize he mic oo ganisms ha ha e in ec ed he
in e io o he plan ma e ial [22]. The disin ec an powe o sodium hypochlo i e depends essen ially on he ac i e
chlo ine con en . Howe e , a e y high pe cen age o chlo ine in a sodium hypochlo i e solu ion would be le hal o
mic ocu s.
5. Conclusion
The disin ec ion echniques used ha e shown ha cashew ee explan s equi e su ace disin ec ion. This can be done
by soaking he explan s in sodium hypochlo i e wi h 1.2% ac i e chlo ine o 2 min. The es ablished p o ocol allowed
he induc ion o one o wo buds pe apex explan om g a ed cashew plan s.
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Compliance wi h e hical s anda ds
Disclosu e o con lic o in e es
No con lic o in e es o be disclosed.
Re e ences
[1] Goujon P., Le eb e A., Le u cq Ph., Ma cellesi A.P. & P alo an J.C. S udies on he cashew ee. F ui s, 28 (3): 217-
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