This article was downloaded by: [The Aga Khan University]On: 09 October 2014, At: 11:02Publisher: Taylor & FrancisInforma Ltd Registered in England and Wales Registered Number: 1072954 Registered office: MortimerHouse, 37-41 Mortimer Street, London W1T 3JH, UK

Caryologia: International Journal of Cytology,Cytosystematics and CytogeneticsPublication details, including instructions for authors and subscription information:http://www.tandfonline.com/loi/tcar20

Application of C-banding in two Arachis wild species,Arachis Pintoi Krapov. and W.C. Gregory and A.villosulicarpa Hoehne to mitotic chromosomeanalysesNeiva I. Pierozzi a , M. Letícia Galgaro b & Catalina R. Lopes ba Centro de Genética, Biologia Molecular e Fitoquímica, Instituto Agronômico deCampinas. Av. Barão de Itapura, 1481. Cx. Postal 28 , Campinas , SP , 13001 -970 , Brasilb Departamento de Genética, IB , Universidade Estadual Paulista , Botucatu , SP , 18603-800 , BrasilPublished online: 28 Jan 2014.

To cite this article: Neiva I. Pierozzi , M. Letícia Galgaro & Catalina R. Lopes (2001) Application of C-banding in twoArachis wild species, Arachis Pintoi Krapov. and W.C. Gregory and A. villosulicarpa Hoehne to mitotic chromosomeanalyses, Caryologia: International Journal of Cytology, Cytosystematics and Cytogenetics, 54:4, 377-384, DOI:10.1080/00087114.2001.10589249

To link to this article: http://dx.doi.org/10.1080/00087114.2001.10589249

PLEASE SCROLL DOWN FOR ARTICLE

Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) containedin the publications on our platform. However, Taylor & Francis, our agents, and our licensors make norepresentations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose ofthe Content. Any opinions and views expressed in this publication are the opinions and views of the authors,and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be reliedupon and should be independently verified with primary sources of information. Taylor and Francis shallnot be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages, and otherliabilities whatsoever or howsoever caused arising directly or indirectly in connection with, in relation to orarising out of the use of the Content.

This article may be used for research, teaching, and private study purposes. Any substantial or systematicreproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in anyform to anyone is expressly forbidden. Terms & Conditions of access and use can be found at http://www.tandfonline.com/page/terms-and-conditions

INTRODUTION

The genus Arachis L. (Leguminosae) is nativeof South America and comprises near 80described annual and perennial species, whichare classified in 9 sections, according toKRAPOVICKAS and GREGORY (1994) and VALLS

(2000).The majority of species is diploid with 2n=20

chromosomes having however three species with2n=18 (A. decora, A. palustris and A. praecox) asobserved by LAVIA (1996, 1998). The occurrenceof tetraploids with 2n=40 is also limited in fourspecies only (A. glabrata, A. hypogaea, A. monti-cola and A. pseudovillosa). Self-pollination is arule in the genus, but seldon some flowers may bevisited by insects which contribute to cross-pol-

lination (KRAPOVICKAS and GREGORY 1994).Although most species reproduce by seeds, someare rhizomatous and reproduce largely throughvegetative means, as A. Pintoi, for instance(KNAUFT and WYNNE 1995), which is alsoemployed as forrage in some tropical countries(VALLS 2000).

The allotetraploid A. hypogaea (peanut) is themost important species in the genus. The culti-vated peanut is considered an important oil seedin developing countries and a valuable source ofprotein for human and animal nutrition (SINGH

and SINGH 1992).Cytological studies in the genus began with

Kawakami’s paper (1930, cited by HUSTED 1931)who counted 40 chromosomes in diploid cells inA. hypogaea. HUSTED (1933, 1936) studied the A.hypogaea chromosome morphology and observedthat they were small. However, he was able todistinguish two conspicuous chromosome pairs:

CARYOLOGIA Vol. 54, no. 4: 377-384, 2001

Application of C-banding in two Arachis wild species,Arachis Pintoi Krapov. and W.C. Gregory andA. villosulicarpa Hoehne to mitoticchromosome analysesNEIVA I. PIEROZZI1, *, M. LETÍCIA GALGARO2 and CATALINA R. LOPES2

1 Centro de Genética, Biologia Molecular e Fitoquímica, Instituto Agronômico de Campinas. Av. Barão de Itapura, 1481. Cx.Postal 28. 13001-970. Campinas, SP, Brasil.

2 Departamento de Genética, IB, Universidade Estadual Paulista, 18603-800. Botucatu, SP, Brasil.

Abstract – Two wild diploid (2n = 20 chromosomes) and self-pollinating Arachisspecies, Arachis Pintoi Krapov and W.C.Gregory and A. villosulicarpa Hoehnewere submmited to C-band technique to karyotype analyses. Root tips wereemployed in the analyses. Morphometric data chose that chromosome lengthsvaried from 3.12 in A. villosulicarpa to 1.45 in A. Pintoi. Karyotype formulaobtained was 10sm to A. Pintoi and 9sm + 1m to A. villosulicarpa. There was apredominance of pericentromeric C-band in all mitotic metaphasic chromo-somes in both species. Besides C-band values, both species still did not differ inrespect to chromosome absolute and relative lengths, centromeric index, sym-metry index and total karyotype haploid length. C-band and morphometric datadid not show strong or significant differences which could separate these twospecies of peanut which belong to evolutive different sections.

Key words: Arachis, C-band, mitotic chromosomes.

* Corresponding author: fax: ++55 019 3242-3602, e-mail:pierozzi@barao.iac.br

Dow

nloa

ded

by [

The

Aga

Kha

n U

nive

rsity

] at

11:

02 0

9 O

ctob

er 2

014

a very small one which was named as “A” and theother pair with a visible secondary constrictionwhich was named as “B” (Satellite - SAT). Later,cytological researches carried out by MENDES

(1947) and CONAGIN (1962, 1963, 1964), estab-lished new chromosome counts as 2n=20 chro-mosomes for wild species.

Studies related to morphological characteri-zations of mitotic chromosomes, in some Arachisspecies, showed a predominance of submetacen-tric chromosomes, based on the arm ratio meanvalues (STALKER and DALMACIO 1981).

Some researchers such as MURTY et al. (1982),KIRTI et al. (1983) or JAHNAVI and MURTY (1985a,b) did studies of chromosome characterizationin some Arachis wild and cultivated species, butchose the meiotic pachytene phase where thechromatin are less condensed. They observed thepresence and distribution of heterochromaticblocks through the chromosomes.

Further, CAI et al. (1987), applied the C-bandtechnique in prometaphasic mitotic chromo-somes of some Arachis species and obtained peri-centromeric C-bands. RAINA and MUKAI (1999)

also observed some pericentromeric heterochro-matin AT-rich bands after DAPI, a selective dye,staining in A. ipaensis and A. villosa.

Preliminary cytological studies has started inour laboratory involving two wild diploid Arachisspecies: A. Pintoi and A. villosulicarpa, with theaim of C-band technique, with the purpose ofchromosome characterization for further com-parative studies. Preliminary results (PIEROZZI etal. 1997) showed that C-band technique is avail-able in these two wild species.

MATERIALS AND METHODS

Plant material

Species identification, provenance and chromo-some numbers are summarized in Table 1.

Pre-treatment, fixation and enzymatic treatment

Seeds of two Arachis species were put to germi-nated in Petri dishes with moisted filter paper at 25°C.

378 PIEROZZI, GALGARO and LOPES

Table 1 – Wild diploid Arachis species studied in this paper:

Section Life form Origin Provenance

* A. Pintoi Krapov. & W.C.Gregory Caulorrhizae perennial ——— CENARGENFernández & Krapovickas (1994)

(2n=20 chromosomes)* A. villosulicarpa Hoehne Extranervosae perennial Mato Grosso state IACMendes (1947)

(2n=20 chromosomes)

CENARGEN - Centro Nacional de Recursos Genéticos. IAC - Instituto Agronômico de Campinas. Chrom Count = chromosomecounting authors.

Table 2 – Chromosome mean values (X) with standard deviation (SD) for absolute length (µm), centromeric index mean val-ues (CI) and chromosome classification (Type) in Arachis Pintoi and A. villosulicarpa.

Arachis Pintoi A. villosulicarpaChrom. Absolute length (µm) CI Type Absolute length (µm) CI Type

X SD X X SD X1 2.98 ± 0.54a 39.85a sm 3.12 ± 0.77a 37.01a sm2 2.80 ± 0.39a 39.36a sm 2.76 ± 0.72a 39.79a sm3 2.66 ± 0.32a 38.42a sm 2.62 ± 0.65a 38.51a sm4 2.49 ± 0.28a 36.33a sm 2.49 ± 0.65a 37.24a sm5 2.42 ± 0.30a 35.48a sm 2.36 ± 0.51a 34.79a sm6 2.31 ± 0.28a 39.49a sm 2.29 ± 0.55a 36.11a sm7 2.12 ± 0.30a 38.68a sm 2.10 ± 0.55a 37.95a sm8 1.98 ± 0.30a 38.80a sm 1.95 ± 0.52a 39.17a sm9 1.75 ± 0.36a 35.72a sm 1.71 ± 0.51a 40.77b m

10 1.45 ± 0.31a 39.36a sm 1.65 ± 0.48a 38.26a sm

Means followed by the same letter = differences were not significant after Tukey test at 1% level.Means followed by different letter = difference was significant after Tukey test at 1% level.

Dow

nloa

ded

by [

The

Aga

Kha

n U

nive

rsity

] at

11:

02 0

9 O

ctob

er 2

014

Roots with approximately 1.5cm of length were colect-ed, treated with a para-dichlorobenzene (p-DB) satu-rated solution for 3 hours at 16ºC, fixed in 3:1 Carnoysolution (100% ethanol and 100% acetic acid, respec-tively) and stored at -20ºC. Fixed roots were treated,for two hours at 32ºC, with pectinase 20% and cellu-lase 2% enzymes which were mixed in equal partsbefore using, for tissue softening.

C-band procedure

Softened roots were placed in 45% acetic acidsolution for 30 minutes at 50°C and squashed. Slideswere placed briefly in liquid nitrogen for the sili-conized coverslips separation with a razor blad, brieflywashed in 100% ethanol, dried and stored for matu-rity (nearly 2 to 3 weeks).

It was employed C-band technique described byMARKS and SCHWEIZER (1974) with some modifica-tions done according to PIEROZZI and JUNG-MEN-DAÇOLLI (1997) suitable for plants with small chro-mosomes, ommiting the brief acid bath after bariumhydroxide incubation.

Chromosome measures

It was calculated for both wild Arachis species themean value and standard deviation for the (1) chro-mosome absolute lengths (µm); (2) chromosome rela-tive lengths (%); (3) total haploid karyotype lengths(µm); (4) total heterochromatin (C-band) expressed aspercentage in relation to the total length of haploid set;(5) C-band percentage in each chromosome in relationto total chromosome haploid length; (6) C-band per-centage in each chromosome in relation to each chro-mosome length; and (7) symmetry index (TF%). Thesymmetry index for each Arachis species was calcu-lated according to HUZIWARA formula (1956, 1962)

named TF% (total form per cent) which is a ratio inpercentage of the total sum of short arm-lengths to thetotal sum of chromosome length. It was also calculat-ed the centromeric index for each chromosome ofboth Arachis species according to GUERRA et al. (1986)which is a complementation of LEVAN et al. (1964)concept.

The mean values of chromosome length (µm and%), of C-band percentage in each chromosome inrelation to total chromosome haploid length and ofcentromeric index were compared by Tukey test afterthe F test. Total haploid karyotype length, total hete-rochromatin C-band and symmetry index mean values(TF%) were compared by t-test (GOMES 1985).

Ten best C-banded spreaded metaphase cells ofeach species were selected for chromosome measuresand for drawings. The drawings were obtained withthe aid of a monocular camera lucida on a Zeiss micro-scope. The schematization was done from the draw-ings with the aid of a dividers.

Photomicrographies were taken using KodakImagelink film, ASA 25, in Vanox-Olympus pho-tomicroscope.

RESULTS

Chromosome absolute length mean valuesvaried from 2.98 µm (chromosome 1) to 1.45µm(chromosome 10) in A. Pintoi and from 3.12 µm(chromosome 1) to 1.65 µm (chromosome 10) inA. villosulicarpa and the differences in chromo-some absolute length were not significant at 1%level after Tukey test, comparing the same chro-mosome in each species. All chromosomes ofA. Pintoi were larger than A. villosulicarpa,excepting chromosomes 1 and 10 which werelarger in A. villosulicarpa than in A. Pintoi (Table2). Both species did not differ through chromo-

C-BANDING IN TWO ARACHIS WILD SPECIES 379

Table 3 – Chromosome mean values (X) with standard deviation (SD) for chromosome relative lehght (%) in Arachis Pintoiand A. villosulicarpa.

Arachis Pintoi A. villosulicarpaChrom. Relative length (%) Relative length (%)

X SD X SD1 12.95 ± 0.82a 13.56 ± 1.46a

2 12.19 ± 0.50a 11.94 ± 0.82a

3 11.58 ± 1.05a 11.38 ± 0.72a

4 10.87 ± 0.59a 10.77 ± 0.63a

5 10.56 ± 0.49a 10.26 ± 0.38a

6 10.09 ± 0.41a 9.95 ± 0.51a

7 9.24 ± 0.77a 9.10 ± 0.39a

8 8.62 ± 0.72a 8.48 ± 1.01a

9 7.06 ± 0.90a 7.42 ± 1.08a

10 6.29 ± 0.80a 7.14 ± 1.13a

Means followed by the same letter = differences were not significant after Tukey test at 1% level.

Dow

nloa

ded

by [

The

Aga

Kha

n U

nive

rsity

] at

11:

02 0

9 O

ctob

er 2

014

some relative legth (%) after Tukey, which variedfrom 12.95% to 6.29% in A. Pintoi and from13.56% to 7.14% in A. villosulicarpa (Table 3).

Total haploid karyotype length mean valueswere 22.97 µm ± 2.95 for A. Pintoi and 22.02µm ± 4.96 for A. villosulicarpa and this differ-ence was not significant after t-test (P<0.01)(Table 4).

The centromeric index mean values variedfrom 35.48 (chromosome 5) to 39.85 (chromo-some 1) in A. Pintoi and from 34.79 (chromo-some 5) to 40.77 (chromosome 9) in A. villosuli-carpa and the differences between the same chro-mosome in each species were not significant at1% level after Tukey test, except for chromo-some 9 (Table 2).

Karyotype formula was 10sm for A. Pintoiand 9sm + 1m for A. villosulicarpa. Symmetryindex measured by TF% mean values were 38.10± 1.96 for A. Pintoi and 37.68 ± 2.82 for A. vil-losulicarpa and the difference was not significantafter t-test (P<0.01) (Fig. 2 and Table 4).

The results with C-band technique showedthat both Arachis species responded positively toC-banding. It was also observed, in interphasecell nuclei, different numbers of darkly stainedspots which had different sizes (Fig. 1A).

C-bands in prometaphase and metaphasemitotic chromosomes were dark and localizedaround centromere region in almost all chromo-somes of both peanut species as seen in Fig. 1 andin idiograms of Fig. 2.

380 PIEROZZI, GALGARO and LOPES

Fig. 1 – Photomicrographies of somatic chromosomes of Arachis species after C-banding. A. prometaphase and B. metaphaseof A. Pintoi; C. prometaphase and D. metaphase of A. villosulicarpa (X 2165). Arrow = C-banded spots in interphase nuclei.Arrow head = chromosome with two C-bands.

Dow

nloa

ded

by [

The

Aga

Kha

n U

nive

rsity

] at

11:

02 0

9 O

ctob

er 2

014

The mean values obtained for total C-bandheterochromatin expressed as percentage inrelation of total haploid karyotype length was17.82% ± 1.99 for A. Pintoi and 18.88% ± 2.11for A. villosulicarpa. Even though A. villosuli-carpa has a higher values than the other species,the difference was not significant after t-test(P<0.01) (Table 4).

C-band percentage mean values in relationto total haploid karyotype length in each chro-mosome varied from 1.57 (chromosome 9) to2.04 (chromosome 6) in A. Pintoi and from1.61 (chromosome 8) to 2.51 (chromosome 1)in A. villosulicarpa. C-band percentage meanvalues in relation to each chromosome lengthvaried from 14.61 (chromosome 2) to 26.63

(chromosome 10) in A. Pintoi and from 17.72(chromosome 2) to 25.65 (chromosome 10) inA. villosulicarpa and these differences consid-ering the same chromosome in each species,were not significant at 1% level after Tukeytest (Tables 5).

It was observed in some metaphasic cellsthat only one pair of chromosome in bothArachis species showed two bands instead one(Figs. 1B, D).

As a consequence of chromosome small size,it was impossible to determine which chromo-some had the NOR (nucleolar organizer region)after C-banding in both wild diploid peanutspecies (Fig. 2). NOR, in fact, did not staineddifferentially after C-banding.

C-BANDING IN TWO ARACHIS WILD SPECIES 381

Fig. 2 – Idiograms of metaphasic chromosomes of A. Pintoi (A) and A. villosulicarpa (B) after C-banding. C = centromere;black segmentes = C-bands.

Dow

nloa

ded

by [

The

Aga

Kha

n U

nive

rsity

] at

11:

02 0

9 O

ctob

er 2

014

DISCUSSION

Chromosome counts for A. villosulicarpa wasfirst established by MENDES (1947) and for A.Pintoi by FERNÁNDEZ and KRAPOVICKAS (1994) as2n=20 chromosomes which were also confirmedin this paper. Chromosomes are small in size, notexcceding 3.2mm of length. None of both speciespresented “A” chromosome, a characteristic ofArachis section, which was first noted by FER-NÁNDEZ and KRAPOVICKAS (1994) and after byLAVIA (1996).

Total haploid karyotype length mean value forA. Pintoi obtained in this paper (22.97µm ± 2.95)was similar to the value obtained byFERNÁNDEZ and KRAPOVICKAS (1994)(23.37µm for haploid set) after Feulgen stainingprocedure. This suggest that the C-band method-ology did not distort in a significant way chro-mosome morphology or length as a consequenceof alcaline treatment (barium hydroxide), asobserved by some authors such as SUMNER(1972).

Great similarity between A . Pintoi and A. vil-losulicarpa was also observed considering now

the morphometric data, such as chromosomelength mean values (µm and %), total karyotypelength mean values, centromeric index and sym-metry index (Tables 2 to 4). This fact was notexpected since both wild groundnut speciesbelong, according to KRAPOVICKAS and GREGO-RY (1994), to different, non-related and evolutivedistant sections. Considering VALLS (2000) infor-mations, section Extranervosae, together withErectoides and Trierectoides, are considered themost primitive in the genus, which gave rise tothe other sections that possess more asymetricalkaryotypes.

Regarding that A. villosulicarpa species is con-sidered taxonomically more primitive than A. Pin-toi, it would be expected to find a more simmetri-cal karyotype formula to the former species. Theanalysis in the karyotype formula (Table 4) showedthat there is exclusively submetacentric chromo-somes in both species, except in A. villosulicarpawhich differed by the presence of one metacentricpair of chromosome. This difference (a tendencyof a more simmetrical karyotype in A. villosuli-carpa), however, was not statistically significant inboth wild peanut species after TF% (Table 4).

382 PIEROZZI, GALGARO and LOPES

Table 4 – Karyotype formula (KF) and mean values (X) with standard deviation (SD) to total haploid karyotype length[THKL(µm)], and symmetry index (TF%) for Arachis Pintoi and A. villosulicarpa.

KF THKL (µm) TF%X SD X SD

Arachis Pintoi 10 sm 22.97µm ± 2.95 ns 38.10 ± 1.96 ns

A. villosulicarpa 9 sm + 1m 22.02µm ± 4.96 ns 37.68 ± 2.82 ns

Ns = non significant after t-test (P< 0.01)

Table 5 – C-band mean values and standard deviation for each chromosome of A. Pintoi and A. villosulicarpa, expressed aspercentage in relation to chromosome length and to total chromosome haploid length, and total heterochromatin C-band meanvalues (%) for each species.

Chrom. C-band / chromosome length (%) C-band / total haploid length (%)Arachis Pintoi A. villosulicarpa Arachis Pintoi A. villosulicarpa

Chrom. X SD X SD X SD X SD

1 15.20 ± 2.3a 19.05 ± 4.9a 2.02 ± 0.37a 2.51 ± 0.74a

2 14.61 ± 2.5a 17.72 ± 5.0a 1.76 ± 0.28a 2.16 ± 0.54a

3 17.09 ± 3.1a 19.33 ± 6.0a 1.74 ± 1.01a 2.20 ± 0.92a

4 16.65 ± 2.7a 20.18 ± 2.9a 1.79 ± 0.25a 2.19 ± 1.04a

5 16.46 ± 3.5a 18.67 ± 4.3a 1.73 ± 0.37a 1.92 ± 0.76a

6 20.25 ± 4.8a 24.45 ± 4.7a 2.04 ± 0.51a 2.43 ± 0.52a

7 18.64 ± 3.7a 20.41 ± 4.6a 1.68 ± 0.34a 1.83 ± 0.41a

8 19.89 ± 4.0a 20.97 ± 5.0a 1.71 ± 0.43a 1.61 ± 0.88a

9 21.35 ± 4.4a 23.23 ± 3.2a 1.57 ± 0.29a 1.71 ± 0.29a

10 26.63 ± 3.2a 25.65 ± 4.0a 1.67 ± 0.24a 1.76 ± 0.44a

Total heterochromatin C-band percentage in relation to chromosome haploid set: Arachis Pintoi: 17.82% ± 2.11 ns; A. villosulicarpa18.88% ± 1.99 ns

Means followed by the same letter = differences were not significant after Tukey test at 1% levelNS = non significative after t-test (P< 0.01)

Dow

nloa

ded

by [

The

Aga

Kha

n U

nive

rsity

] at

11:

02 0

9 O

ctob

er 2

014

Even though A. Pintoi and A. villosulicarpabelong to different and non related sections theystill presented the same tendency of only peri-centromeric C-band formation (Fig. 2) whichindividuals values, expressed as percentage, didnot differ neither in relation to chromosomelength (C-band/chromosome length) nor to hap-loid karyotype length (C-band/total haploidlength) (Table 5). CAI et al. (1987) working witheight different Arachis species from sectionsArachis and Erectoides also observed a highdegree of similarities among them in relation toC-band occurence and position, almost all werelocalized around the centromere and few of themwere terminal and intercalary. The appearenceof these two last types of C-band may be due tomitotic phase chosen, prometaphase, in whichchromosomes are less condensed thanmetaphase, allowing better visualization of inter-calary and/or terminal C-bands. Another possi-bility for the presence of intercalary C-band is theacid treatment step (HCl before barium hydrox-ide solution) as observed by CARVALHO andSARAIVA (1993) in maize and which was not pre-sent in our experiment.

According to GALASSO et al. (1993) similarC-banded patterns may be expected in phyloge-netic correlated species and this also reflects sim-ilarities in relation to heterochromatin composi-tion or organization.

Some similarities in relation to C-banded kary-otypes showing preferencial pericentromericC-bands in some other genera of Leguminosaefamily with small mitotic chromosomes (less than4 µm of length) were also noted by some authors,such as LAVANIA and SHARMA (1980) in somespecies of Lathyrus, GALASSO et al. (1992 and1993) in some species of Vigna, or as TAYYAR etal. (1994) in some annual Cicer species.

Practically, no heterozigozity for C-band pat-tern was observed in both species of Arachis. Thismigth be a consequence of self-pollinating natureof Arachis species. This fact was also observed byTAYYAR et al. (1993), in some species of anotherleguminous self-pollinating genera, Cicer.

In relation to A. villosulicarpa, it seems thatthe pericentromeric C-bands observed in thispaper may be related with those pericentromer-ic heterochromatic blocks observed by JAHNAVI

and MURTY (1985a, b) in pachytene meiotic chro-mosomes .

Molecular studies done by GALGARO et al.(1998) in some Arachis species with the aim of

RFLP and RAPD methodology, showed that A.Pintoi differed from A. villosulicarpa in relation toDNA band numbers and positions.

Based on these informations and consideringmorphometric data and C-band pattern, it is pos-sible that during the evolutive process whichinvolved A . Pintoi (sect. Caulorrhizae) and A . vil-losulicarpa (sect. Extranervosae), the chromosomelength, the centromeric position, the heterochro-matin quantity and position did not change great-ly in a way that could be seen by C-band method-ology. Changes might be occurred at genic level,as could be seen by GALGARO et al. (1998).

However, more studies in other Arachisspecies, involving not only perennial but alsoannual too, with the aim of C-band techniquehave to be done to know more about hete-rochromatin distribution in the genus.

Acknowledgements – We thank BL JandimarC.Farhat for English review, Ms. Antonia A.M.Fer-reira for technical works, Drs. A. S. Pompeu,I. Godoy and Renato F.A.Veiga (IAC) for seed sup-plies.

REFERENCES

CAI Q., LU S. and CHINNAPPA C.C., 1987 – Analy-sis of karyotypes and Giemsa C-banding patternsin eight species of Arachis. Genome, 29: 187-194.

CARVALHO C.R. and SARAIVA L.S., 1993 – A newheterochromatin banding pattern revealed bymodified HKG banding technique in maize chro-mosomes. Heredity, 70: 515-519.

CONAGIN C.H.T.M., 1962 – Espécies selvagens dogênero Arachis. Observações sobre os exemplaresda coleção da S. de Citologia. Bragantia, 21: 341-358.

–, 1963 – Número de cromossomos das espécies sel-vagens de Arachis. Bragantia, 22: 125-129.

–, 1964 – Número de cromossomos em Arachis sel-vagem. Bragantia, 23: 25-27.

FERNÁNDEZ A. and KAPROVICKAS A., 1994 – Chro-mosomas y evolucion em Arachis (Leguminosae).Bonplandia, 8: 187-220.

GALASSO I., PIGNONE D. and PERRINO P., 1992 –Cytotaxonomical studies in Vigna. I. Generaltechnique and Vigna unguiculata C-banding.Caryologia, 45: 155-161.

–, 1993 – Cytotaxonomic studies in Vigna. II. Hete-rochromatin characterization in Vigna unguicula-ta and three related wild species. Caryologia, 46:275-282.

C-BANDING IN TWO ARACHIS WILD SPECIES 383

Dow

nloa

ded

by [

The

Aga

Kha

n U

nive

rsity

] at

11:

02 0

9 O

ctob

er 2

014

GALGARO L., LOPES C.R., GIMENES M., VALLSJ.F.M. and KOCHERT, G., 1998 – Genetic varia-tion between species of sections Extranervosae,Caulorrhizae, Heteranthae and Triseminatae(genus Arachi) estimated by DNA polymorphism.Genome, 41: 445-454.

GOMES F.P., 1985 – Curso de estatistica experimen-tal. Livraria Nobel Editora, Piracicaba.

GUERRA M.S., 1986 – Revewing the chromosomenomenclature ol Leva et al. Braz. J. Genet., 9:741-743.

HUSTED L., 1931 – Chromosome number in speciesof peanut, Arachis. Amer. Naturalist, 65: 476-477.

–, 1933 – Cytological studies on the peanut, Arachis.I. Chromosome number and morphology. Cytolo-gia, 5: 109-117.

–, 1936 – Cytological studies on the peanuts, Arachis.II. Chromosome number, morphology and behav-ior, and their application to the problem of the ori-gin of the cultivated forms. Cytologia, 7: 396-423.

HUZIWARA Y., 1956 – Karyotype analusis in somegenera of Compositae . I. Karyotype of JapaneseEupatorium. Cytologia, 21: 114-123.

–, 1962 – Karyotype analysis in some genera of Com-positae. VIII. Further studies on the chromosomesof Aster. Amer. J. Botany, 49: 116-119.

JAHNAVI M.R. and MURTY U.R., 1985a – Chromo-some morphology in species of the Sections Erec-toides and Extranervosae of the genus Arachis L.Cytologia, 50: 747-758.

–, 1985b – A preliminary pachytene analysis of twospecies of Arachis L. Theor. Appl. Genet., 70:157-165.

KIRTI P.B., BHARATHI M., MURTY U.R. and RAON.G.P., 1983 – Chromosome morphology in threediploid species of Arachis and its bearing on thegenome of groundnut (Arachis hypogaea L.).Cytologia, 48: 139-151.

KNAUFT D.A. and WYNNE J.C., 1995 – Peanutbreeding and genetics. Advances in Agronomy,55: 393-445.

KRAPOVICKAS A. and GREGORY W.C., 1994 – Tax-omonia del genero Arachis (Leguminosae). Bon-plandia, 8: 1-186.

LAVANIA U.C. and SHARMA A.K., 1980 – GiemsaC-banding in Lathyrus L. Bot. Gaz., 14: 199-203.

LAVIA G.I., 1996 – Estudios cromosómicos enArachis (Leguminosae). Bonplandia, 9: 111-120.

–, 1998. Karyotype of Arachis palustris and A. prae-cox (Section Arachis). Two species with basic chro-mosome number x=9. Cytologia, 63: 177-181.

LEVAN A., FREDGA K. and SANDBERG A.A., 1964 –Nomenclature for centromeric position on chro-mosomes. Hereditas, 52: 210-220.

MARKS G.E. and SCHWEIZER D., 1974 – Giemsabanding karyotype differences in some species ofAnemone and Hepatica nobilis. Chromosoma,44: 405-421.

MENDES A.J.T., 1947 – Estudos citológicos no gêneroArachis. Bragantia, 7: 257-267.

MURTY U.R., KIRTI P.B., BHARATHI M. and RAON.G.P., 1982 – Identification of the chromosomesof groundnut Arachis hypogaea L. Cytologia, 47:585-594.

PIEROZZI N.I. and JUNG-MENDAÇOLLI S.L., 1997 –Karyotype and C-band analysis in two Genipa L.species (Rubiaceae, Gardenieae tribe). Cytologia,62: 81-90.

PIEROZZI N.I., GALGARO M.L. and LOPES C.R.,1997 – Estudos de caracterização cromossômicaem algumas espécies de Arachis. Resultados pre-liminares. In: I Simpósio Latino-Americano deRecursos Genéticos Vegetais. Campinas, Brasil,pp. 40.

RAINA S.N. and MUKAI Y., 1999 – Genomic in situhybridization in Arachis (Fabaceae) identifiesthe diploid wild progenitors of cultivated(A. hypogaea) and related wild (A. monticola)peanut species. Plant Syst. Evol., 214: 251-262.

SINGH U. and SINGH Y., 1992 – Tropical grainlegumes as important human foods. EconomicBotany, 46: 310-321.

STALKER H.T. and DALMACIO R.D., 1981 – Chro-mosomes of Arachis species, section Arachis. J.Hered., 72: 403-408.

SUMNER A.T., 1972 – A simple technique for demon-stration centromeric heterochromatin. Exp. CellRes., 75: 304-306.

TAYYAR R.I., LUKASZEWSKI A. J. and WAINES J.,1993 – Chromosome banding patterns in theannual species of Cicer. Genome, 37: 656-663.

VALLS J.F.M., 2000 – Diversidade genética no gêneroArachis e a origem do amendoim. In: Anais do17° Encontro Sobre Temas de Genética e Mel-horamento, pp. 19-33. Piracicaba, Brasil.

Received July 27, 2001; accepted September 26, 2001

384 PIEROZZI, GALGARO and LOPES

Dow

nloa

ded

by [

The

Aga

Kha

n U

nive

rsity

] at

11:

02 0

9 O

ctob

er 2

014