Pakistan J. Agric. Res. Vol 24 No. 1-4, 2011. EVALUATION OF KASURI METHI TRIGONELLA FOENUM- GRAECUM L.VAR. TO ESTABLISH GI RIGHT OF PAKISTAN Shazia Erum, Rashid Anwar and Shahid Masood* ABSTRACT:- Geographical indicator (GI) of crops create a positive impression of the product quality, the environmental virtue and human skill of the area. In the present study, comparative analysis of total seed proteins of Kasuri methi (GI of Kasur, Pakistan) was evaluated with other Trigonella genotypes by SDS-PAGE. Results showed that at protein level Kasuri methi acquired a unique status as a G.I of Kasur. Cluster analysis (UPGMA) of 28 genotypes including both methi and methray from various agro ecological zones of Pakistan were interlinked to some extent however Kasuri methi make their identity by standing alone among tested genotypes on the dendrogram, due to the geographical environment of Kasur in Punjab. In this connection ‘Benefit sharing’ mechanism can be accelerated by escalating farmer’s participation for more cultivation in the interest of country. Key Words: Trigonella foeum-graecum; Seeds; Diversity; Geographical Indicator; Phenotype; Pakistan. INTRODUCTION industries, etc. Pharmaceutically medici- Fenugreek, Trigonella foenum-graecum nal species of the genus Trigonella are T. L. is an ancient and annual legume crop foenum-graecum L., T. balansae, T. mainly grown for multiple uses in many corniculata, T. maritima, T. spicata, T. parts of the world. Trigonella, comes from occulta,T. polycerata, T. calliceras, T. cretica, Latin meaning ‘little triangle’, refers to the T. caerulea, T. lilacina, T. radiata, T. spinosa triangular shape of its small yellowish- (Petropoulos, 2002; Basu, 2006). The bio- white flowers. The species epithet foenum- logical and pharmacological actions of graecum means ‘Greek hay’ (Rosengarten, fenugreek are attributed to the variety of 1969). It is also called ‘ox horn’ or ‘goat horn’ its constituent, namely, steroids, N-com- because of the two seed pods projecting in pounds, polyphenolic substances, volatile opposite directions usually from the nodes constituents, amino acids, etc. of the stem base that resemble ox or goat (Mehrafarin et al., 2010). horns (Petropoulos, 2002). Landraces and Fenugreek seed contains 45-60% car- species of Trigonella have been found in bohydrates, mainly mucilaginous fiber parts of Europe, northern Africa, west and (galactomannans), 20-30% proteins high in south Asia, north and south America and lysine and tryptophan, 5 - 10% fixed oils Australia (Basu, 2006; Acharya et al., (lipids), pyridine alkaloids, mainly trigonel- 2006). line (0.2 - 0.38%), choline (0.5%), Fenugreek leaves and seeds are con- gentianine and carpaine, the flavonoids sumed in different countries around the apigenin, luteolin, orientin, quercetin, world for different purposes such as medi- vitexin and isovitexin, free amino acids, cine (anti-diabetic, lowering blood sugar and such as 4-hydroxyisoleucine (0.09%), argi- cholesterol level, anti-cancer, anti-micro- nine, histidine and lysine, calcium and bial, etc.), making food (stew with rice in iron, saponins (0.6 - 1.7%), glycosides yield- Iran, flavor cheese in Switzerland, syrup, ing steroidal sapogenins on hydrolysis (di- mixed seed powder with flour for making osgenin, yamogenin, tigogenin, flat bread in Egypt, curries, dyes, young neotigogenin), cholesterol and sitosterol, seedlings eaten as a vegetable, etc.), roasted vitamins A, B1, C and nicotinic acid and grain as coffee-substitute (in Africa), con- 0.015% volatile oils ‘n-alkanes’ and ‘ses- trolling insects in grain storages, perfume quiterpenes’ (Budavari, 1996; Newall et al., *Institute of Agricultural Biotechnology and Genetic Resources Institute, National Agricultural Research Cen- tre, Islamabad, Pakistan.. 25 SHAZIA ERUM ET AL. 1996; Mehrafarin et al., 2010). attributed as a major reason for failure to Most renowned Pakistani fenugreek consider the possibility that farmers have with remarkable aroma cultivated in the an intellectual investment in their folk Kasur district of Punjab province well varieties (Cleveland and Stephen, 1997; known as ‘Kasuri methi’. Kasur (total area Nagarajan, 2005). 3,995 km2, elevation 218 m.a.s.l). Methi The objective of the study is to evalu- (fenugreek) from Kasur is very famous for ate the Trigonella species collected from its fragrance throughout the country and various agro ecological zones of Pakistan became geographical indicator of Kasur as and to distinguish Kasuri methi at protein Kasuri Methi, so it can be claimed/patent level by SDS- PAGE. for GI or AO. ‘Appellation of Origin’ (AO) means that a product originates in a spe- MATERIALS AND METHODS cific geographic region and the character- The research was conducted at Na- istic qualities of the product are due to the tional Agricultural Research Centre, geographical environment, including natu- Islamabad, Pakistan. Seeds of 28 genotypes ral and human factors (Nagarajan, 2005). of Trigonella (Trigonella foenum graecum- The farming community contributions Methi and Trigonella corniculata-Methray) have to be recognized and rewarded and were collected from various agroecological benefit sharing should become mandatory. zones of Pakistan including Mianwali, Relationship between farmers’ variety (FV) Pakpattan, Bahawalpur, Faisalabad, and GI provides certain rights (Plant Pro- Shorkot, Narowal, Karachi, Okara, tection Variety and Farmers Rights Act Talwandi, Bahawalnagar, Daska, Sargodha, 2001 in India) to farmers, such as to save, Quetta and Kasur according to the quad- use, sow, re-sow, exchange, share or sell rate method. The current study was initi- his farm produce including that of the reg- ated to investigate the potential electro- istered variety. In Pakistan, farmers who phoresis for intra specific characterization develop new plant varieties like any other of Trigonella on the basis of their total seed plant breeder can apply their material for protein and explore distinctiveness of the conduct of Distinctness, Uniformity and kasuri methi (landrace) with other geno- Stability (DUS) testing and registration. In types of Trigonella. the last hundred years there has been a For judgment of Kasuri methi as a G.I., drive for improved agriculture and that has seeds samples were divided into two groups. replaced farmers’ variety in several crops Group I consists of 22 seed samples of with new varieties developed by the plant Trigonella foenum graecum-Methi and Group breeders. Yet farmer’s variety is still domi- II comprises seed samples of Trigonella nant in pulses, vegetables, melons, etc. corniculata-Methray, along with Kasuri There are lots of controversial patent cases methi seed sample in both the groups. involving traditional Knowledge and Ge- Genetic diversity of Trigonella species netic Resources including hoodia cactus, evaluated by sodium dodecylsulphate poly- turmeric, neem, ayahuasca and basmati rice acrylamide gel electrophoresis (SDS-PAGE) case, facing problems for their patent rights according to the standard method as geographical indicators. The GI for agri- (Leammli, 1970; Rabbani et al., 2001; cultural goods like Basmati rice, coffee, tea, Ahmed et al., 2009 and 2010; Turi et al., wine, etc. revolve around consumer pref- 2010; Cheema et al., 2010). The subunit erences for the palate feeling, aroma and molecular weight of the protein bands was physical appearance that enhances the determined according to Weber and Osborne appetite. An ideal mixture of all these at- (1969). Each band was considered as a tributes raises the value of the product due character for which the presence or ab- to reasons of consumer preference sence was coded 1 or 0 respectively in a (Nagarajan, 2005). Lack of documentation data matrix for numerical analysis. Den- of indigenous farmers’ plant breeding is sity of the protein band was not considered. 26 EVALUATION OF KASURI METHI Unweighted pair group method with an major groups were observed on the dendro- arthimetic average (UPGMA) cluster analy- gram (Figure1), where first and third group sis was used to infer genetic relationships showed 100% similarity among themselves and phylogeny among the genotypes of (including 1,2,3,4,5,19,18,17,16,15,14,6,7,8 Trigonella. All computations were done by and 20,21) while 23% deviation was ob- using the NTSYS-pc, Version 2.2 package served in the second group (9,13,11,10 and (Rohlf, 2005; Rabbani et al., 2001 and 12) whereas the genotype No. 22, popularly 2008). For an effective breeding programme, known as “Kasuri Methi” from Kasur (22) information regarding the extent and na- stand alone and expressed itself as a G.I ture of genetic diversity within a crop spe- even at protein level. cies is essential. It is particularly very use- The variable banding pattern shown by ful for characterization of individual acces- Kasuri methi made it different from other sions and as a guide in the selection of par- genotypes. As proteins are the translational ents for hybridization programme. products of genes therefore, the differences in proteins could be related with such dif- RESULTS AND DISCUSSION ferences in genes responsible for synthe- Seed protein analysis by SDS-PAGE has proved to be an effective way of reveal- Table 1. Passport data of Trigonella ing the differences and relationship be- germplasm tween taxa. The high stability of the seed Accession Town/city protein profile and its additive nature make Group I seed protein electrophoresis a powerful tool PAK 020978 Mainwali in elucidating the origin and the evolution