CYTOTAXONOMY

OBSERVATION :

The investigator tried to study the cytotaxonomy of nematodes

together with their morphology. During the course of cytotaxonomy it was

found that the best stage of karyometric analysis appears to be the pachytene

stage prepared from oogonial cells. The division of which start at third larval

stage and continue upto early adult stages. This division is normal mitotic

division and results into large number of oogonia having somatic 2n

chromosomes. The reason behind not selecting other stages of cell division is

basically the karyometric analysis is not possible on other stages because cell

stages were full of errors due to the presence of small and fine chromosome

which could not be avoided. A systematic list of nematode parasites studied

during investigation are as under :-

Family: Oxyuridae Cobbold, 1864.

Genus: Mirzaiella Basir, 1942

M. meerutensis n. sp.

Genus: Chitwoodiella Basir, 1948

C. asiatica n. sp.

Family: Aoruridae Skrjabin and Schikobalova, 1951.

Sub family: Aorurinae Walton, 1927.

Genus: Hammerschmidtiella Chitwood, 1932

H. indicus n. sp.
Sub family: Blattophilinae Skrjabin and Schikobalova 1951.

Genus: Leidynema Schwenk, 1929.

L. orientalis n. sp.

Family: Thelastomatidae Travassos, 1929

Sub family: Thelastomatinae Travassos, 1929

Genus: Thelastoma Leidy, 1849

T. alii Farooqui, 1970

Genus: Schwenkiella Basir, 1956

S. longicaudata (Meyer, 1896) Basir, 1956

S. icemi (Schwerk, 1926) Basir, 1956

S. orientalis Singh and Agarwal, 1997

Genus: Gryllophila Basir, 1942

G. basiri n. sp.

Genus: Binema Travassos, 1925

B. ornate Travassos, 1925

B. atrophicaudata n. sp.

Genus: Isobinema Rao, 1958.

I. jairajpurii n. sp.

Genus: Psilocephala Rao, 1958

P. gryllotalpae n. sp.

As regard the number of chromosomes are concerned in all those

nematodes it is 2n = 10. All these nematodes have two types of chromosome
submetacentric and metacentric of varying numbers except Schwenkiella.
The Schwenkiella has three different type of chromosomes viz.,
submetacentric, metacentric and subtelocentric. The detailed observation of
pachytene karyometry appended in Table 23- 34 and Fig 1-28).
As regard the total chromatin length is concerned it was found to be highest in

Leidynema orientalis n. sp. 0.057 mm and lowest in S. orientalis Singh and

Agarwal, 1997, 0.214. However, with in the genus Schwenkiella it range

between 0.0214 - 0.038 mm and in Binema 0.0392 - 0.0401 mm.

S% was found to be highest in Binema atrophicaudata n.sp.

(61.05%) and lowest in S. icemi (25.17%). No pecularity of this value was

noted in different species of the same genus for their taxonomic significance.

The TF% was found to be highest in S. icemi (48.44%) and lowest in H.

indicus (26.92%). This value exhibit generic peculiarity but when comparison

of this value was made with the different species of the same genus no

conclusion could be drawn.

The average length of the chromosomes also exhibits generic

characteristics. The largest chromosome length was found in L. orientalis

(0.0114 mm) and smallest in S. orientalis (0.00428 mm). The study of

average long arm length from the taxonomy point of view, it was found that

their exists inter generic peculiarity of this parameter. It was found to be

highest in L. orientalis and lowest in S. orientalis being 0.0072 mm and

0.002752 mm respectively.

Average short arm length also exhibits generic peculiarity like long

arm length, being highest in L. orientalis 0.00352 mm and lowest in S.

orientalis 0.001466 mm. The study of average arm ratio shows both inter

genric and inter specific peculiarity of this parameter. It was found to be

highest in L. orientalis (2.19%) and lowest in T. alii (1.592%).

 The critical evaluation of average centromeric index exhibits inter

generic peculiarity being highest in B. ornata (39.04%) and lowest in L.

orientalis (30.346%).

The study of average TCL% also exhibit its taxonomic value within

different generabeing highest for C. asiatica (25.386%) and lowest for T. alii

(199 93%).

With the help of comparative karyometric analysis and reliable inter

specific difference between the relative lengths of different chromosomes in

all the five pairs have been traced. Essential inter specific differences were

noted in most of the parameters like-. Total chromatin length, S%, TF%,

average long and short arm length, F%, TCL% and average centromere

indices. The karyotype of these nematodes has been studied for the first time.

It is interesting to note that all the five pairs of chromosomes in nematodes

under observation are provided with well developed short arms.

The detail study of centromeric indices of different nematodes and

subjecting it to UPGMA analysis. It was found that Binema and Mirzaiella are

closely related with each other showing clusturing index at 38.5795. Similarly

Schwenkiella and Thelastoma are closely related showing C.I. 36.957. In all

the ten nematode can be separated in two group showing C.I at 34.5195.

First group includes Schwenkiella, Thelastoma, Isobinema,

Hammerschmidtiella and Psilocephala showing C. I. at 36.957, 36.3595,

35.8118 and 35.2848 respectively.

 However, the other group includes Leidynema (C.I. = 37.9584),

Gryllophila (C. I . = 38.3099), Chitwoodiella (38.5238), Binema and Mirzaiella

at 38.5795.

DISCUSSION
Cytogenetic work with nematodes was initiated in the later part of the

nineteenth century and continued with high intensity upto the second decade

of twentieth century. This early work was motivated by the urge to understand

and explain the role of chromosomes in cell division, reproduction and

heredity. The animal parasitic nematodes especially Parascaris equorum,

provided very favourable cytological material Triantaphyllou (1983). The work

of Von Beneden (1883) for the first time, demonstrated, the process of

meiosis involving the reduction of chromosomes from diploid to haptoid stage

during gametogenesis and the re-establishment of diploid number during

fertilization. Meanwhile, Boveri (1887) reported chromatin diminution in the

somatic cells during embryogenesis of ascarid nematode P. equorum.

Similarly, existence of compound chromosomes was first observed in

maturing ocytes of Parascaris. The chromosomal basis of sex determination

was another cytological feature which was studied extensively during that

early period. The gynogenesis as modified method of reproduction, was

described first of all in nematodes after very elegant experimental work of

Kruger (1913).

Following the early outburst of cytological discoveries, investigations

on nematode cytology and cytogenetics proceeded very slowly and have

maintained almost same slow pace till recently. In the last few decades,

interest in the nematode cytogenetics has been renewed, but only for

economically important groups such as plant and animal parasitic nematodes.

The work of Triantaphyllou (1983) brought about a revolution in

chromosome cytology of nematodes and other helminths. However, the

progress in study of chromosomes in nematodes, which appears to form the

intermediate step in the invertebrate evolutionary ladder thus, holds the key to

understand the inverterbrate phylogeny, also remained very slow. The

available chromosomal data on nematodes in general including insect

parasitic nematodes pertain only to two hundred different species, though

there are estimates of the existence of 0.5 million species of nematodes all

over the world in different biotopes capable of supporting life.

With the application of newer techniques the primary difficulty, of

obtaining good mitotic figures, could not be solved. The study of cytology is

predominently performed on gonads, particularly ovary, because of being

hologonic in nature. Therefore, there is a good scope for chromosomal

analysis in economically important species of nematodes.

Though, most of the chomosomal investigators on nematodes

concentrated their study on oogenesis but the advantages of chromosomal

data are many like -

1. Cytotaxonomy
2. Karyotype study and

3. Evolutionary studies etc.

In nematodes, the use of chromosomal informations as a tool in

taxonomy has even an added importance since, nematode classification is

beset with many difficulties.

Moreover, since this group is originator of many other phyla of

Eedysozoa like Acanthocephala, Nematomorpha, etc. The karyotypic data in

them can help in understanding the evolutionary pathways of these groups.

Moreover, it can also tell us that how this group of animals diversified in such

a way to inhabit all sorts of biotopes and occur in large number in a wide

variety of shapes, sizes and structures. The study of chromosomes of

nematodes thus, may give an insight into these events.

Some groups of nematodes, specially parasites of vegetables have

been found to have extensive chromosomal polymorphism (Goswami, 1979).

Their chromosomal analysis can therefore be useful in understanding the

process of speciation. Besides this, it can also be used in deciphering the

sibling and cryptic species (Patterson and Stome, 1952). Moreover, study of

chromosome can also reveal the secret of nature behind the evolutionary

diversity of this compact group of inverterbrates by virtue of which they are

successfully adapting themselves in the changing environmental conditions

with a constant pace.

 Chromosome number, an important feature of nematode karyotype, is

useful in interpreting relationships among the species of a genus, or genera of

a family. As regards the number of chromosomes are concerned, the data

varies from n = 1, as in Diploscapter coronata (Hatchler, 1968), to n = 19, as

in plant parasitic nematode Anguina tritici (Triantaphyllou and Hirschmann,

1966). Higher chromosome numbers are also encountered in polypoid forms

of nematodes, like - Ditylenchus (n = 24). Meloidogyne (3n = 54) etc. During

the course of study the chromosome number in the Thelastomatoidea was

found to be five pairs only. However, there were variations about the number

of chromosomes. As regard the type of chromosomes are concerned, mostly

they were submetacentric and metacentric except genus Schwenkiella, where

the chromosomes were subtelocentric also, besides, being metacentric and

submetacentric. However, it is difficult for the author to correlate her findings

because previously no such attempt has been made by the workers, for the

reasons best known to them. But, I am confident that this parameter can

successfully be deployed to work out the taxonomy of nematodes.

Total chromatin length S%, average length of chromosomes, average

short arm length, centromeric indices, TCL% etc. also exhibit significant

intergeneric and inter specific variations. Thus, these parameters can be

successfully used as a powerful cytogenetic tool to resolve taxonomic

confusions in the nematode systematics as it is widely used in other animals

and plants by workers like Mc Clintock (1929), Barton (1950), Gillies (1968),
Ramanna and Wagenvoort (1976), Dundas et. al. (1983), Khus et al. (1984),

Singh and Hymowitz (1991) and Kumar and Gupta (1997).

Presence of well developed short arms proves that all these

nematodes have evolved recently or they are not very primitive in origin

(White, 1973).

The UPGMA analysis reveals the evolutionary hierarchy of ten different

genera of nematodes under observation. But it is difficult for me to

corroborate these findings as no such attempt has been made in past.