KARYOMORPHOLOGICAL AND STOMATAL STUDIES OFALLIUM HOOKERITHWAITES

AlliumhookeriThwaites belongs toAmaryllidaceae family is one of the most important medicinal plants, enormously used in cancer or inflammation because it contains large amounts of sulfonylmethane.It is also used to treat coughs, colds, fatigue and to recover immunity. Karyotypic study showed that somatic chromosome number is 2n = 22. Only nearly sub median and nearly median chromosomes are found in the complement. Karyotype formula = 1 nsm(-)+ 10 nm=2n=22. The total length of long arms is 60.36 Ã‚Âµm, ranging from 3.16Ã‚Âµm to 8.23Ã‚Âµm. Total length of short arms is 31.57Ã‚Âµm, ranging from 1.75Ã‚Âµm to 4.11Ã‚Âµm. The total length of the chromosome is 92.17Ã‚Âµm, ranging from 4.78Ã‚Âµm to 12.35Ã‚Âµm. Relative chromosome length ranges from38.70Ã‚Âµm to 100 Ã‚Âµm. Arm ratio ranges from1.70 to2.38.Tf %= 41.63.Centromeric index ranges from 0.40 to 0.28 and The total chromatin index is 100, ranging from 5.18 to 13.40. Anomocytic type of stomata is observedStomatal index and stomatal size were calculated. The maximum length (53.6Ã‚Â±2.06Ã‚Âµm) was observed in basal portion of the adaxial surface and the minimum length (45.2 Ã‚Â± 1.40Ã‚Âµm) was observed in the middle portion of abaxial surface. The maximum width size (24.8Ã‚Â±1.11Ã‚Âµm) was observed in the basal portion of the adaxial surface and the minimum width was (18.8 Ã‚Â± 1.14Ã‚Âµm) at the middle portion of the abaxial surface. Stomatal index 12.63Ã‚Â±0.5) maximum and minimum (6.13Ã‚Â±0.40) was observed.


…………………………………………………………………………………………………….... Introduction:-
The study ofchromosomes are considered as an important tool for determining the evolutionary trend phylogeny and systematic position of the related species (Sharma and Sharma, 1959;; Watanabe et al ., 1995;Das et al., 1999;Vanzel et al., 2000;Shan et al., 2003; Sanjaykumar and Tsipila Thonger.,2018). Chromosomes are considered as the most important component of the genetic system of the eukaryotes. For better understanding of any species chromosome number, structure and behavior is determined qualitatively and quantitatively (Naruhashi and Iwatsubo,1991). The chromosomal study are quite helpful in genetic improvement of the crops whether the crops are rare, endangered or threatened species. The research on chromosome provides the basic foundation of the entire discipline of the molecular genetics of the present day world..Allium hookeri belongs to family amaryllidaceae (Revised by APG system in 2016 i.e.4 th classification till now). Its common name is hooker's chive and is also known as winter leek. Locally it is called as ' Van (Sharma et al.,2011) and is enormously used in cancer or inflammation because it contains large amounts of sulfonyl methane (Bae and Bae , 2012).It is also used to treat coughs, colds, fatigue and to recover immunity. This plant has gained significant attention because it produces high amount of phenolic antioxidants, phytosterols, fibres, ascorbic acid, flavonoids and allicin. The chromosome number recorded were 2n= 22 which is regarded as the most common to the best of our knowledge, there is no report on the karyology of this species in Jharkhand. Therefore the aim of present investigation is to provide information on the mitotic chromosome and stomata to understand the detailed cytotaxonomic position ofAllium hookeriThwaites.

Material and Method:-
The plant of Allium hookeriwas collected from Lalkhatanga , Ranchi, Jharkhand which is 20 km away from the main city. Following studies were done on this plant.

Karyotype study
For karyotypic study the root apices of 1-2 cm were cut and pretreated with saturated PDB for 4 hours and fixed in Carnoy's fluid 1 (3:1 ethanol: glacial acetic acid ) for 24 hours and preserved in 70% ethanol at 4 0 c for future use. The preparation of slide was carried out by hydrolyzing the preserved root tips in 1N Hcl for 10-12 mins and stained in 2% acetocarmine solution (Sharma and Sharma, 1980) and slides wereprepared using squash technique.For analyzing chromosomes a well scattered metaphase stage were selected and observed under compound microscope at a magnification of 10X and 45X and photomicrography was done using DSLR camera.

Karyotype preparation
Following parameters were used for the analysis of karyotype i.e., length of long arm (LA), Short arm(SA) and the Total chromosome length(TL),Total length of long arms (TLLA), Total length of short arms(TLSA) and Total length of the whole chromosomes (TLWC), Relative chromosome length (RCL), Arm Ratio (AR) , Centromeric Index (CI), Chromosome type (CT), Karyotype formula (KF) etc.The chromosomes were classified on various categories following the Kutarekar and Wanjari(1983)classification.Based on the centromeric position the chromosomes are classified into metacentric (M), Submetacentric (Sm) and Subtelocentric (St) having an arm ratio (Long/short) above 0.76, 0.75 to 0.51 and less than 0.50 respectively.On the basis of length the chromosomes were grouped into following categories:-Type A = 11.00µm & above. Type B = 9µm -10.99µ,Type C = 7.00µm -8.99µm, Type D = 5.00µm -6.99µm, Type E = 3.00µm -4.99µm.The other indices were also used to analyse the karyotype asymmetry such as:-

Result:-Karyotype
In this species, the chromosome number was observed to be 2n = 22 chromosomes in which 10 nearly median and 1 nearly sub median chromosomes were observed. The total length of long arms is 60.36 µm, ranging from 3.16µm to 8.23µm. Total length of short arms is 31.57µm, ranging from 1.75µm to 4.11µm. The total length of the chromosome is 92.17µm, ranging from 4.78µm to 12.35µm. Relative chromosome length ranges from38.70µm to 100 µm. Arm ratio ranges from1.70 to2.38.Tf %= 41.63.Centromeric index ranges from 0.40 to 0.28. and The total chromatin index is 100, ranging from 5.18 to 13.40. On the basis of the length, the chromosome were classified into Type A-E (Table: 1 (Table: 2)

Stomata
The stomatalstudies are depicted in table (3) and in fig (3). Stomatal index and stomatal size of AlliumhookeriThwaites were calculated. The measurement of abaxial and adaxial surface of leaves at the apex, middle and base were observed. The maximum length (53.6±2.06µm) was observed in basal portion of the adaxial surface and the minimum length (45.2 ± 1.40µm) was observed in the middle portion ofabaxial surface. The maximum width size (24.8±1.11µm) was observed in the basal portion of the adaxial surface and the minimum 130 width was (18.8 ± 1.14µm) at the middle portion of the abaxial surface. Stomatal index 12.63±0.59 which was maximum in the adaxial surface and minimum in the apex portion of theabaxial surface (6.13±0.40).