CHEMICAL METHODS FOR VARIETAL IDENTIFICATION = A large number of crop improvement programmes are running and with the result of this a large number of varieties are being developed. " Thus varietal identification becomes an essential issue to maintain the genetic purity and identity of each variety. Further, the aspect of Distinctness, Uniformity and Stability (DUS) is fundamental for characterization of varieties. Accurate identification of varieties is not only a pre requisite for DUS testing, but is critical for the production of quality seed also. * Maintenance of genetic purity of varieties is of primary importance for preventing varietal deterioration during successive regeneration cycles and for ensuring varietal performance at an expected level. * Laboratory tests have several additional benefits for varietal identification. These chemical tests are very quick, easy to do, reproducible and can be conducted throughout the year under controlled conditions.Priacipte of chemioal mvethod * APALE Hon Morphotagloal descriptors, there by a need to screen for atiitional descriptors with significant discriminating power and vepradieiiility, that are not lafluenced by the environment, Seeds coloue revetion 0 different chemloals is one such alternative, Whew seeds are exposed to certaly chemicals, they develop a eharacteriatic colour depending on its chemical or metabolite eonstitation, This color is a variety specific trait and the application of this concept for the purpose of varietal characterlmation and Wentitication hax now been well established ln different crops, * Hor example, modified phenol test ln wheat, KON test li ree, FeSO, teat i foxtall inilet, peartuillet and sorghum,Types of Chemical tests Phenol test Peroxide test KOH Bleach test NaOH test FeSO, test GA; soaking test Alkaloid test lodine testpe . Phenol Test Used for varietal identification in crops viz., Wheat, rice, pearl millet, sorghum, foxtail millet etc. Itis recommended by ISTA (1999). It is easy, quick and reliable test. The test is highly specific and monogenically controlled by a dominant gene. The principle behind the phenol test is that the mono or di or poly phenol oxidized by the enzyme tyrosinase present in pericarp /mesocarp/any other seed structure uses phenol as substrate. As a result of this enzymatic oxidation activity dark coloured insoluble melanin pigment is formed. Colour reaction serves as a basis for grouping. The intensity of colour developed is evaluated on a 0 — 9 selected scale (0 — negative or no change in colour and gradual positive intensification of colour from light brown to deep black graded from 1 — 9).. Peroxidase test 15 Soybean seeds are placed in test tubes and add 10 drops of 0.5 % guiacol. After 10 min, a drop of 0.1 % hydrogen peroxide Peroxidase is added. +ve or high peroxidase activity showed a reddish brown solution whereas, peroxidase -ve or low peroxidase activity showed colourless solution.. KOH bleach Test The presence or absence of tannic acid (dark pigment in the testa or under coat layer of sorghum seed). It is mainly done for sorghum to distinguished dark & light pigmented testa. Seeds are soaked in 1:5 ( KOH:Naocl) for 5 to 10 min then seeds are dried to observe the color. Based on colour reaction varieties are identified whether single/ pure OR an admixture of different types. Deep wine red color (+ve) and No staining show (-ve) reaction.4. NaOH Test wo It is used to differentiate in the colour of seeds of garden and fodder peas. Seeds are soaked in 1% NaOH Solution (for 1 hrs). Colour changed is classified as - dark yellow, light yellow & no colour. Reasons for NaOH & KOH Colour change is mainly due to inherent chemical difference and secondary metabolites present in the seeds. . Ferrous sulphate (FeSO,) test Seeds soaked in 1.5% FeSO, for 4 hrs under ambient conditions then taken out. After removal of excess moisture, seeds are evaluation colour change and grouped as: dark grey streaks, Brown streaks, Brown spots on the husk of cereal seeds.6. GA; soak test = Seedlings raised in germination paper towels moistened with GA; solution (100 ppm). « After 7-10 days, seedling growth is observed and cultivars are grouped on the response to shoot & root elongation as high & low. = For example, gibberelic acid test used for varietal identification of cotton genotypes, grouped into low response & very low response genotypes. i7. Alkaloid test " In Lupinus species, the Presence or absence of alkaloid is a diagnostic feature. The seeds are soaked in water for 24 hours. Next, a thin slice from each seed is cut and over a white surface. Subsequently, one or two drops of Lugol’s solution are added. = Adistinct brown-red precipitate indicates the Presence of alkaloid. placed on a glass plate 8. lodine test " Differentiate millet grain seeds from weed seeds. = After soaking the seeds in 5% iodine solution for 5 to 7 minutes the weed seeds develop dark color and millet seeds remain same or changed into a light pink color.Conclusion = Combination of different chemical tests is suggested. = A single chemical test is not sufficient to identify varieties, though they are simple and efficient but not sufficient. ® In combination with each other, they can separate almost any number of varieties and thus, have immense value for the varietal identification.