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Systematic variation in leaf amino acid compositions of leguminous plants
Phyrochenustry, Vol 23, No 10,PP 2227-2229,1984 Pnntcd m GreatBntam
0031-9422/84s300+000 0 1984Per8amonPtess Ltd
SYSTEMATIC VARIATION IN LEAF AMINO ACID COMPOSITIONS OF LEGUMINOUS PLANTS HOCK-HIN YEOH, YEOW CHIN WEE and LESLIE WATSON* Botany Department, Natlonal Umverslty of Smgapore, Kent Ridge, Singapore 0511, *Taxonomy Unit, Research School of Blologlcal Sciences, The Austrahan NatIonal Umverslty, P 0 Box 475, Canberra, ACT , Austraha (Reutsed recetued 12 March 1984) Key Word Index-Legununosae, leaf ammo aclds, systematic vartatlon
Abstract-The leaf protein content for 17 species of legumes ranges from 2 8 to 9 4g % fr wt, mlth an average of 5 3 g % fr wt Taxonomlc pattern JSdetectable m leaf ammo acid patterns, those of the Mlmosoldeae being dlstmgulshable from those of the Paplhonoldeae and Caesalpmloldeae
INTRODUCTION Analyses of available data on seed ammo actd composittons of legumes have shown vanatlons consistent
with grouping of the Legummosae mto the subfamllles, Mlmosoldeae, Caesalpmloldeae and Paplhonoldeae [l, 21 There JS also evidence of correlation between vmt.l susceptlbdity and taxonomlc groupings m the case of certain legume hosts [3] In dlcotyledonous plants, ammo acid compositions of leaves have yielded taxonomlc dlstmctlons between groupings, e g between caryophyllolds, legumes and acantholds [4] while in a monocotyledonous family, the Poaceae, the existence of taxonomlc pattern m the ammo acid compositions of leaves, caryopses and nbulose-1,5-blsphosphate carboxylase, has also been demonstrated [5-71 As part of our continuing effort to understand the taxonomlc predictablhty of protein ammo acid composltlons of plants, it seemed worthwhile to examme the Legummosae to discover whether taxonomlc pattern can be detected m the leaf amm6 acid composltlons of a dlcotyledonous family, and whether the legume pattern IS dlstmgulshable from that of the grasses [7] This paper presents the ammo acid analyses of leaves from 17 species of legumes, representmg all three subfamlhes [ 81 RESULTSAND DISCUSSION Leaf protein content for the 17 species of legumes studied showed large vanatlon, rangmg from 2 8 g % fr wt m Tamartndus andtca to 9 4 g y0 fr wt m Bauhmra purpurea (Table 1) Compansons at subfinuly level do not show any taxonomlc correlation m the dlstrlbutlon pattern of leaf protein content, the mean protein content in the Mlmosoldeae (5 2g o/0 fr wt), Caesalpmloldeae (5 3 g % fr wt) and Papdlonoldeae (5 6g % fr wt) being closely similar Vanatlons m the leaf ammo acid cornpositIons for the 17 species of legumes are gwen m Table 1 Of these, Munosa pudlca and Bauhmra purpurea are mterestmg m that they have yielded the hqhest level of Leu (I 3.8 %) and Asp (16 0 %) respectively Compatrsons of the three subfarmhes show that the Mimosoideae pattern JS dlstmgmshable from that of the Paplhonoldeae by its
slgmficantly higher (at 5 % probabdlty level) Glu and HJS and lower Asp and Thr, and from the Caesalpmloldeae by its significantly higher Glu and HJSand lower Thr and Phe content (Table 1) The ammo acid profiles of the Caesalpmloldeae and Paplhonoldeae do not show any sign&ant differences at the 5 % probabtity level although some ammo acid values of the Caesalpmloldeae (such as Asp, Glu, Met and His) occupy an intermediate posItJon between the Mlmosoldeae and the Paplhonoldeae Although the sample of species JS small, this result JS consistent with taxonomlc schemes which place the Papdlonoldeae closer to the Caesalpmloldeae than to the Mlmosoldeae [3,8]_ Comparison of the legume leaf ammo acid patterns with those of grasses [7] shows that they are clearly datmgmshabie, the legumes having yielded slgruficantly higher levels of Asp, Pro, Tyr, Phe, HJS and Arg and lower levels of Ser and Ala These observations seem to support the suggestion that a systematic survey of a wide range of plant groups may reveal predictable leaf protein ammo acid compositions [4, 71
EXPERIMENTAL Hunt materrnlsLeafsamples were collected from plants grown tn the garden of the Botany Department, National Untverslty of Smgapore, the Singapore Botanic Gardens and elsewhere m Smgapore Their IdentItles were checked wth reference to remonal floras and voucher specunens have been deposited In the National Umverslty of Singapore Botany Department
Herbanum Preparatwnjorammoacid analysrsLeaf blades of mature and healthy leaves were finely cut and MO-2CNlmg matenal was hydrolysed m 0 5 ml 3 N mercaptoethanol sulphotuc acid m a sealed tube at 110” for 22 hr [7] After hydrolyses, 0 5 ml 2 N NaOH was added to the sample which was then diluted with 2 ml &t&d water The sample (100 4) was then analysed on a Be&man ammo acid analyser 119CL Duphcate analyses rep resentmg collections from tierent plants of the same species camed out for some sptcles showed some vanations (see Table 1) Such vmtlons among tndlvlduals of the same smes have been observed m our earher studies [7] but so far they have not a&ted agndicantly the taxononuc concluslons. For subsequent 2227
Taxonomlc mean value
Taxonormc mean value Papdlonotdeae Baphla nrtufa Afzel Crotalarza retusa L Desmadtum trzforum (L ) DC Mtllettla atropurpurea Benth Psophocarpus tetragonolobus (L Psophocarpus tetragonolobus (L Pterocarpus u&us Wdld Sesbama grandzflora Pers
) DC ) DC
Taxononuc mean value Caesalplmotdeae Bauhuua purpurea L Cassla blpora Gnseb Cassla blpora tiseb Delonzx regza Ralin Saraca tha~pmgensls Cantley ex Pram Tamarmdus rndlca L
Mlmosotdeae Acacza aunculrfomus A Cunn ex Benth Acacia aurwdtiormls A Cunn ex Benth Adenanthera pavonrna L Alblzurfalcatarta (L ) Fosberg Munosa pudzca L Parkta Javamca (Lamk ) Merr
132
55
53 55 53 51 58 57 59 55
122 140 134 124 124 121 129 153
53 57 55 61 49 51
160 127 122 128 125 118 54
50
121
131
51 48 52 48 49 49
Thr
121 120 119 119 126 121
Asp
54
59 49 52 59 51 51 60 48
54
48 56 52 59 52 56
54
63 58 52 57 47 51
Ser
129
124 134 131 130 123 125 119 141
123
119 131 125 134 108 127
140
150 150 130 146 134 139
Glu
65
76 58 59 77 56 58 74 54
66
60 59 59 68 76 68
66
63 68 61 74 57 70
55
53 57 55 57 54 54 53 54
54
65 56 58 37 56 56
60
51 54 59 53 71 66
61
61 61 59 60 66 67 59 57
62
51 62 68 68 62 62
59
58 58 63 60 58 57
49
47 51 54 50 49 47 49 47
47
48 48 48 48 43 41
48
46 46 52 45 47 49
18
17 18 18 18 18 20 15 17
19
16 21 22 21 17 20
20
20 21 19 19 21 18
36
34 36 37 38 37 35 38 34
35
35 37 36 35 33 36
35
30 33 37 34 33 37
97
109 92 91 89 110 114 93 91
96
105 90 100 86 96 96
100
89 88 92 92 138 92
52
52 50 52 50 55 54 56 52
54
55 53 52 52 61 51
51
45 49 56 49 48 54
61
60 59 62 59 62 61 68 61
64
63 61 65 63 68 61
58
54 58 63 58 54 63
Ammo md composltlons (g % total ammo aclds) Pro Gly Ala Val Met Ile Leu Tyr Phe
Table 1 Total leaf ammo acid composlhons of legumes
25
26 28 27 22 23 25 25 21
24
24 23 24 26 24 24
28
30 31 29 26 25 28
His
66
62 60 71 75 66 65 63 65
71
53 69 67 67 90 80
65
80 73 66 75 44 62
Lys
02
01 03 00 00 01 02 01 04
02
03 01 01 01 01 02
00
01 00 00 01 00 00
Trp
43
44 47 45 41 47 46 39 48
45
44 49 46 47 40 45
46
43 45 48 45 48 44
hg
5 57
631 309 580 424 660 674 647 642
530
943 542 5 32 565 332 275
5 16
359 308 745 446 702 3 52
(g% fr vJt)
Total ammo actds
e
m
z
j: 4
X
Systematic variation m the ammo acid cornposItIons of LRgummous plants taxonomlc mean calculations mean values of duplicate analyses were used Total lea! ammo aclds Total leaf ammo acid contents were calculated from the ammo acid analyses, and expressed as g y0 fr wt leaf samples Acknowledgements-We thank Kath Brett for performmg the ammo acid analyses Tins work IS supported m part by research grant (RP 83/81) from the NatIonal Umverslty of Smgapore REFERENCES
1 VanEtten, C H , Miller, R W , Wolff, I A and Jones, Q (1963)
2229
Agrzc Food Chem 11,399 2 VanEtten, C H , Kwolek, W F, Peters, J E and Barclay, A S (1967) Agrz Food Chem 15, 1077 3 Gibbs, A and Watson, L (1980) Advances In Legume Saence (Summerfield, R J and Bunting, A H ,eds)Vol 1, p 239 Brat Mus Nat HIS, England 4 Watson, L and Creaser, E H (1975) PhytocItemlstry 14,121l 5 Yeoh, H H and Watson, L (1981) Phytochemrstry 20, 1041 6 Yeoh, H H and Watson, L (1982) Phytochmnrstry 21, 71 7 Yeoh, H H and Watson, L (1982) Phytochemtstry 21, 615 8 Polhdl, R M and Raven, P H (1981) Advances WI Legume Systemutxs Royal Botanic Gardens, Kew Crown
0031-9422/84s300+000 0 1984Per8amonPtess Ltd
SYSTEMATIC VARIATION IN LEAF AMINO ACID COMPOSITIONS OF LEGUMINOUS PLANTS HOCK-HIN YEOH, YEOW CHIN WEE and LESLIE WATSON* Botany Department, Natlonal Umverslty of Smgapore, Kent Ridge, Singapore 0511, *Taxonomy Unit, Research School of Blologlcal Sciences, The Austrahan NatIonal Umverslty, P 0 Box 475, Canberra, ACT , Austraha (Reutsed recetued 12 March 1984) Key Word Index-Legununosae, leaf ammo aclds, systematic vartatlon
Abstract-The leaf protein content for 17 species of legumes ranges from 2 8 to 9 4g % fr wt, mlth an average of 5 3 g % fr wt Taxonomlc pattern JSdetectable m leaf ammo acid patterns, those of the Mlmosoldeae being dlstmgulshable from those of the Paplhonoldeae and Caesalpmloldeae
INTRODUCTION Analyses of available data on seed ammo actd composittons of legumes have shown vanatlons consistent
with grouping of the Legummosae mto the subfamllles, Mlmosoldeae, Caesalpmloldeae and Paplhonoldeae [l, 21 There JS also evidence of correlation between vmt.l susceptlbdity and taxonomlc groupings m the case of certain legume hosts [3] In dlcotyledonous plants, ammo acid compositions of leaves have yielded taxonomlc dlstmctlons between groupings, e g between caryophyllolds, legumes and acantholds [4] while in a monocotyledonous family, the Poaceae, the existence of taxonomlc pattern m the ammo acid compositions of leaves, caryopses and nbulose-1,5-blsphosphate carboxylase, has also been demonstrated [5-71 As part of our continuing effort to understand the taxonomlc predictablhty of protein ammo acid composltlons of plants, it seemed worthwhile to examme the Legummosae to discover whether taxonomlc pattern can be detected m the leaf amm6 acid composltlons of a dlcotyledonous family, and whether the legume pattern IS dlstmgulshable from that of the grasses [7] This paper presents the ammo acid analyses of leaves from 17 species of legumes, representmg all three subfamlhes [ 81 RESULTSAND DISCUSSION Leaf protein content for the 17 species of legumes studied showed large vanatlon, rangmg from 2 8 g % fr wt m Tamartndus andtca to 9 4 g y0 fr wt m Bauhmra purpurea (Table 1) Compansons at subfinuly level do not show any taxonomlc correlation m the dlstrlbutlon pattern of leaf protein content, the mean protein content in the Mlmosoldeae (5 2g o/0 fr wt), Caesalpmloldeae (5 3 g % fr wt) and Papdlonoldeae (5 6g % fr wt) being closely similar Vanatlons m the leaf ammo acid cornpositIons for the 17 species of legumes are gwen m Table 1 Of these, Munosa pudlca and Bauhmra purpurea are mterestmg m that they have yielded the hqhest level of Leu (I 3.8 %) and Asp (16 0 %) respectively Compatrsons of the three subfarmhes show that the Mimosoideae pattern JS dlstmgmshable from that of the Paplhonoldeae by its
slgmficantly higher (at 5 % probabdlty level) Glu and HJS and lower Asp and Thr, and from the Caesalpmloldeae by its significantly higher Glu and HJSand lower Thr and Phe content (Table 1) The ammo acid profiles of the Caesalpmloldeae and Paplhonoldeae do not show any sign&ant differences at the 5 % probabtity level although some ammo acid values of the Caesalpmloldeae (such as Asp, Glu, Met and His) occupy an intermediate posItJon between the Mlmosoldeae and the Paplhonoldeae Although the sample of species JS small, this result JS consistent with taxonomlc schemes which place the Papdlonoldeae closer to the Caesalpmloldeae than to the Mlmosoldeae [3,8]_ Comparison of the legume leaf ammo acid patterns with those of grasses [7] shows that they are clearly datmgmshabie, the legumes having yielded slgruficantly higher levels of Asp, Pro, Tyr, Phe, HJS and Arg and lower levels of Ser and Ala These observations seem to support the suggestion that a systematic survey of a wide range of plant groups may reveal predictable leaf protein ammo acid compositions [4, 71
EXPERIMENTAL Hunt materrnlsLeafsamples were collected from plants grown tn the garden of the Botany Department, National Untverslty of Smgapore, the Singapore Botanic Gardens and elsewhere m Smgapore Their IdentItles were checked wth reference to remonal floras and voucher specunens have been deposited In the National Umverslty of Singapore Botany Department
Herbanum Preparatwnjorammoacid analysrsLeaf blades of mature and healthy leaves were finely cut and MO-2CNlmg matenal was hydrolysed m 0 5 ml 3 N mercaptoethanol sulphotuc acid m a sealed tube at 110” for 22 hr [7] After hydrolyses, 0 5 ml 2 N NaOH was added to the sample which was then diluted with 2 ml &t&d water The sample (100 4) was then analysed on a Be&man ammo acid analyser 119CL Duphcate analyses rep resentmg collections from tierent plants of the same species camed out for some sptcles showed some vanations (see Table 1) Such vmtlons among tndlvlduals of the same smes have been observed m our earher studies [7] but so far they have not a&ted agndicantly the taxononuc concluslons. For subsequent 2227
Taxonomlc mean value
Taxonormc mean value Papdlonotdeae Baphla nrtufa Afzel Crotalarza retusa L Desmadtum trzforum (L ) DC Mtllettla atropurpurea Benth Psophocarpus tetragonolobus (L Psophocarpus tetragonolobus (L Pterocarpus u&us Wdld Sesbama grandzflora Pers
) DC ) DC
Taxononuc mean value Caesalplmotdeae Bauhuua purpurea L Cassla blpora Gnseb Cassla blpora tiseb Delonzx regza Ralin Saraca tha~pmgensls Cantley ex Pram Tamarmdus rndlca L
Mlmosotdeae Acacza aunculrfomus A Cunn ex Benth Acacia aurwdtiormls A Cunn ex Benth Adenanthera pavonrna L Alblzurfalcatarta (L ) Fosberg Munosa pudzca L Parkta Javamca (Lamk ) Merr
132
55
53 55 53 51 58 57 59 55
122 140 134 124 124 121 129 153
53 57 55 61 49 51
160 127 122 128 125 118 54
50
121
131
51 48 52 48 49 49
Thr
121 120 119 119 126 121
Asp
54
59 49 52 59 51 51 60 48
54
48 56 52 59 52 56
54
63 58 52 57 47 51
Ser
129
124 134 131 130 123 125 119 141
123
119 131 125 134 108 127
140
150 150 130 146 134 139
Glu
65
76 58 59 77 56 58 74 54
66
60 59 59 68 76 68
66
63 68 61 74 57 70
55
53 57 55 57 54 54 53 54
54
65 56 58 37 56 56
60
51 54 59 53 71 66
61
61 61 59 60 66 67 59 57
62
51 62 68 68 62 62
59
58 58 63 60 58 57
49
47 51 54 50 49 47 49 47
47
48 48 48 48 43 41
48
46 46 52 45 47 49
18
17 18 18 18 18 20 15 17
19
16 21 22 21 17 20
20
20 21 19 19 21 18
36
34 36 37 38 37 35 38 34
35
35 37 36 35 33 36
35
30 33 37 34 33 37
97
109 92 91 89 110 114 93 91
96
105 90 100 86 96 96
100
89 88 92 92 138 92
52
52 50 52 50 55 54 56 52
54
55 53 52 52 61 51
51
45 49 56 49 48 54
61
60 59 62 59 62 61 68 61
64
63 61 65 63 68 61
58
54 58 63 58 54 63
Ammo md composltlons (g % total ammo aclds) Pro Gly Ala Val Met Ile Leu Tyr Phe
Table 1 Total leaf ammo acid composlhons of legumes
25
26 28 27 22 23 25 25 21
24
24 23 24 26 24 24
28
30 31 29 26 25 28
His
66
62 60 71 75 66 65 63 65
71
53 69 67 67 90 80
65
80 73 66 75 44 62
Lys
02
01 03 00 00 01 02 01 04
02
03 01 01 01 01 02
00
01 00 00 01 00 00
Trp
43
44 47 45 41 47 46 39 48
45
44 49 46 47 40 45
46
43 45 48 45 48 44
hg
5 57
631 309 580 424 660 674 647 642
530
943 542 5 32 565 332 275
5 16
359 308 745 446 702 3 52
(g% fr vJt)
Total ammo actds
e
m
z
j: 4
X
Systematic variation m the ammo acid cornposItIons of LRgummous plants taxonomlc mean calculations mean values of duplicate analyses were used Total lea! ammo aclds Total leaf ammo acid contents were calculated from the ammo acid analyses, and expressed as g y0 fr wt leaf samples Acknowledgements-We thank Kath Brett for performmg the ammo acid analyses Tins work IS supported m part by research grant (RP 83/81) from the NatIonal Umverslty of Smgapore REFERENCES
1 VanEtten, C H , Miller, R W , Wolff, I A and Jones, Q (1963)
2229
Agrzc Food Chem 11,399 2 VanEtten, C H , Kwolek, W F, Peters, J E and Barclay, A S (1967) Agrz Food Chem 15, 1077 3 Gibbs, A and Watson, L (1980) Advances In Legume Saence (Summerfield, R J and Bunting, A H ,eds)Vol 1, p 239 Brat Mus Nat HIS, England 4 Watson, L and Creaser, E H (1975) PhytocItemlstry 14,121l 5 Yeoh, H H and Watson, L (1981) Phytochemrstry 20, 1041 6 Yeoh, H H and Watson, L (1982) Phytochmnrstry 21, 71 7 Yeoh, H H and Watson, L (1982) Phytochemtstry 21, 615 8 Polhdl, R M and Raven, P H (1981) Advances WI Legume Systemutxs Royal Botanic Gardens, Kew Crown