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Organization and Classification of

Cytochrome P450 Genes in Castor (Ricinus
communis L.)
Maryada Shailendar Kumar, Peram
Ravindra Babu, Khareedu Venkateswara
Rao & Vudem Dashavantha Reddy
Proceedings of the National
Academy of Sciences, India Section B:
Biological Sciences
ISSN 0369-8211
Volume 84
Number 1
Proc. Natl. Acad. Sci., India, Sect. B Biol.
Sci. (2014) 84:131-143
DOI 10.1007/s40011-013-0192-8

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Rao  V.) Maryada Shailendar Kumar • Peram Ravindra Babu • Khareedu Venkateswara Rao • Vudem Dashavantha Reddy Received: 3 December 2012 / Revised: 13 April 2013 / Accepted: 7 May 2013 / Published online: 12 June 2013 Ó The National Academy of Sciences.Author's personal copy Proc.com Introduction Castor (Ricinus communis L) is an important non-edible oil seed crop belonging to the family Euphorbiaceae.1007/s40011-013-0192-8 RESEARCH ARTICLE Organization and Classification of Cytochrome P450 Genes in Castor (Ricinus communis L. Distinctive phylogenetic groups of castor CYPs showed varying levels of conserved gene organization. High level (*90 %) of ricinoleic acid present in castor oil contributes to the stable viscosity index and high lubricity even under low temperature conditions and hence castor oil is used in the manufacture of antifreeze fuels and lubricants for space rockets. among the subfamily members conserved sequences as well as length of exons and phasing of introns have been observed. terpenoids.  Exon–intron phasing  Arabidopsis M. the EEC and Japan. In general. Keywords Cytochrome P450  Ricinus communis L. manufacture of a wide range of products like nylon fibre. CYP702. Cytochrome P450s represent *1 % of plant proteome and constitutes one of the largest family of enzymes controlling primary and secondary metabolism. Castor bean oil besides its use as vegetable and medicinal oil has several industrial applications. Castor is predominantly cultivated in India. Hyderabad 500007. India 2013 Abstract Castor is an important non-edible oilseed crop with several industrial applications. hydraulic fluids. Natl. Sci. CYPs in various plants are known to perform reactions of both primary and secondary metabolism and are involved in the production of fatty acids. India e-mail: vdreddycpmb@yahoo. V. Sci. USSR.. CYP736 and CYP749) are not present in the castor genome. Kumar  P. CYP729. Dehydrated castor oil is used in the paint and varnish industry. China. Cytochrome P450s(CYPs) represents a large class of enzymes which mediate diverse metabolic reactions in almost all organisms such as bacteria. India. The genus Ricinus is monotypic with R. Babu  K. Castor is diploid (2n = 20) and is presumed to be a secondary-balanced polyploid with basic number of chromosomes x = 5 [2]. Maximum number (92) of CYP450 genes possessed single intron followed by intron less genes(35). B Biol.two intron containing genes (25) and four intron containing genes (20). However. artificial leather. jet engine lubricants. Cytochrome P450s have been found to play vital role in the metabolism and detoxification [7]. Sect. sterols. The crop is extensively grown in tropics and sub-tropics as well as temperate regions. bullet proof glass and bone prostheses [3–5]. D. Cytochrome P450s catalyze oxidation of various substrates using oxygen and NAD (P) H [8. CYP708. CYP705. CYP728. plant hormones. A novel gene RcCYPN could be identified in the present study. variable intron length(s) recorded was attributed to continuous genome expansion. Brazil. these 123 . animals and plants. (Jan–Mar 2014) 84(1):131–143 DOI 10. Furthermore. Reddy (&) Centre for Plant Molecular Biology. S. 9]. India accounts for nearly 60 % castor production [3]. plastics. Based on sequence similarity with Arabidopsis orthologs and CYP nomenclature these genes have been classified into 45 families representing 77 subfamilies and grouped into ten clans. Deduced CYP proteins of castor on an average exhibited 485 amino acid residues. R.000 genes [6]. The sequence analysis established genome size of the castor as *320 Mb with an estimated number of 31. fibre optics. flavonoids. Genes pertaining to ten CYP families (CYP80. Acad. lignin and signalling molecules etc. CYP92. communis as the only species [1]. Osmania University. fungi. Analysis of castor genomic resources identified 210 putative Cytochrome P450 genes. CYP733.

if any. CYP712 and distributed into 33 subfamilies(Fig. 7 with seven introns. CYP94. Of these. phytozome.net/) was searched for putative Cytochrome P450 genes and retrieved the predicted protein and nucleotide (CDs and Genomic) sequences. and CYP86 are multi-family clans. all single intron containing CYP genes (84) are grouped into A-type families. CYP77. CYP94. an equal number of introns represent phase one (88) and phase two (80) organization. Only one of the 13 genes in the CYP82 family is intron less and 11 genes are with single intron. of which 104 are with phase zero followed by 18 with phase one and 8 with phase 2.0 database (http://www. The NeighbourJoining (NJ) tree method by P-distance in MEGA5 [13] was used to construct the phylogenetic tree. CYP706. Of the 92 single intron containing genes majority (87) of them exhibited phase 0 introns. CYP716. The clan CYP72 containing seven families viz.313 bp and a mean of 385 bp were observed. A total of 495 introns were recorded in all 175 intron containing CYP genes. Among the genes (83) with multiple introns. CYP721. Naming of castor CYP genes was carried out based on similarity with the gene orthologs of Arabidopsis. The clan 86 with CYP86. 1). 5 with three introns. In view of economic importance of castor and the central role of CYPs in the plant metabolism. CYP78. CYP77. The non-A type CYP genes are distributed into 27 families consisting of 44 subfamilies (Fig. The chemical diversity across plant species is well correlated with the diversity of CYPs. CYP703. CYP97. CYP707. Among the 210 CYP genes. Alignment of CYP coding sequences to genomic sequences using BLAT program was carried out predicting the number of introns. S. CYP701. CYP72. CYP710. On the other hand non-A-type genes (87) contained 365 introns of which 223 with phase zero. Material and Methods Castor genome at phytozome v 8.000 bp upstream of 50 end. Results Sequence analysis of castor genome disclosed 210 putative CYP coding genes grouped into ten clans (CYP51. The significance level for the NJ analysis of phylogenetic tree using bootstrap testing with 1. exonic lengths and total length of intronic sequences.CYP75. However the intron length varied from 26 to 12. Kumar et al. CYP83. Among the intron containing genes a total of 670 exons with varied length from 9 to 1. Use of the generated alignments computed the lengths of introns. 74. six clans are single family clans. CYP88. CYP722. CYP76. CYP72. com/berry. 70 with phase 1 and 72 with phase 2 (Table 1). CYP74. 71. CYP85.ac. and CYP704 families contain 21 genes.9 and gap extension penalty 0). BLASTP analysis of castor proteome with retrieved CYP protein orthologs was carried out to identify diversified paralogs. The CYP proteins which are below 300 and above 600 amino acids were validated by using Softberry gene prediction tool (http://linux1. CYP93. CYP720. CYP81. While. and CYP724 families. Except for 8. CYP734. CYP96 and CYP98 families.217 bp with an average of 328 bp. 85. CYP79. CYP73. Multiple sequence alignment of castor CYP proteins was performed using the UPGMB clustering (Gap opening -2. CYP98. About 44 % castor CYP genes are with single intron. CYP715. 710. 97.ebi. The alignment of predicted amino acid sequences of CYPs from the castor with genomic DNA sequence using Wise2 program (http://www. in the MUSCLE module [12] from the Mega5 software. 86.000 replications was carried out. and single genes each with 13 and 15 introns. CYP87. 2). CYP89. respectively. CYP79. .711 and 727) consisting of 45 families represented by 77 subfamilies (Table 1). if any. CYP84. CYP51. CYP714. CYP718. remaining three clans. All the introns showed canonical GT–AG splice sites. The non-A type CYP genes (87) are distributed in the other nine clans. CYP82. CYP96. 18 families. CYP709. Out of them 327 introns exhibited phase zero.uk/Wise2/) was carried out to identify positions of introns and exons and their phases. 25 genes are with two introns followed by 20 with four introns. Maximum number (92) of CYP genes are with single intron.. Maximum number of ten intron less genes are seen in the family CYP89 followed by eight genes in the family CYP94. CYP86. possessed 20 genes. CYP89. CYP711 and CYP727. While. The clan CYP85 comprising of 36 genes represent CYP85. CYP84. CYP90. besides identifying novel CYP coding genes. followed by four genes with phase 1 introns and one gene with phase 2 intron. 14 with eight introns. 175 genes are split genes and the remaining 35 genes are intron less and are present in CYP74. CYPs are also responsible for the degradation of endogenous and exogenous compounds which are harmful to plants [11]. CYP82. 3 each with five and six introns. The clan CYP727 present in castor is absent in the Arabidopsis genome. The CYP genes in castor code for proteins in the range of 298–632 amino acids with an average of 485 residues.phtml) by increasing the scaffold size to 2. 4 with nine introns. 72. A-type CYP genes (123) possessed 130 introns.Author's personal copy 132 proteins represent *1 % protein coding genes of any given organism [10].softberry. the present study has been mainly focused on structural analysis of CYP genes from castor and their classification. and CYP735. The highest number (123) of the CYP genes are present in the CYP71 clan which represents the whole set of A-type CYP genes belonging to 123 M. CYP71.

m000806 CYP85 clan RcCYP74A1 30170.2.376. 352. 82.245.0 0 Intron (s) phase 7. 216. 84.1.0.0.182 1. 78. 88.356 606. 194 91.0. 226 1.507 3.0. 116.1.m013972 CYP74 clan RcCYP72A14 29633.340 286.256.432 259.221.245.135 2. 26 461.m000932 RcCYP51G1 CYP gene name 29739.0. 101 89.391. 126.370 910 217. 267. 88 96.m003138 30076.0 1.364.429 169. 290. 274 2.245. 94. 278 1.88.379. 122.2.m003779 29983.687 910 2.m014292 RcCYP85A1 29790.376.096 1. 5.245. 87. 210 343.968 2.426 280.0.644 3.2.996 2.426 462.379.2.429 295.79.160 112. 369 286.516 2.379.426 280. 82.0.m008568 CYP51 clan Castor gene ID 8.0 1. 186. 265.540 2.796 3.291 4.0.581 626 582 1.072 2.2.0.0.196 2. 102 114.0. 112.m009066 30174. 110.2.0. 92.797 2.251. 46. 564.0. 157 159.m004472 29794.126.0. 110.874 0 4.063.423 103. 58.95. 172.0. 92.2. 217.m004534 29848.426 295.251.2 2.2.0. 72.379.221.0 0.224.493. 232. 92.227.m009065 30174.165 2. 227.2.999 Exon(s) size (bp) 84.233.0 1.0 1. 182. 153. 84 80.0.0. 91 0 4.376.0 1.0.0.0 1. 75 402 Intron (s) size (bp) 2.0.0.79.533 2.0 1.426 277.1. 77 99.218.1. of amino acids Author's personal copy Organization and Classification of Cytochrome P450 133 123 . 364. 101.181.322.87.527 0 1.265. 166.254.379.251 471.233.2. of Intron (s) 197.221.m001078 30174.m009068 30205. 151.0.252.0.107.0 1.224.159.233.2 0.417.385. 578.206.140 79.0.173. 79.115 98. 77.105 1.m000415 30910. 141. 99. 248.453 492.233.349.m000878 RcCYP87A5 RcCYP87A2 30190.57 104. 90.2.0.93.128.88 1.m009070 30174.150.150.245.985 590 837 734 333 375 1.919 1.125.0. 95.942 4.0 0.356 566 432 389 555 522 402 Sum of intron length (bp) 473 487 544 435 416 496 518 515 514 452 475 516 513 529 613 343 445 512 493 512 523 503 509 488 509 413 510 486 No.0. 100. 235 1.322.81.701 2.118 1.m000018 RcCYP87A4 27985.246. 121.426 301.m009067 30174.0.0.150. 116.224. 632.429 94.414 14.0.2. 88.0.0 1.218.0.1.m011135 30147.0.128.0. 101. 131. 219. 89.0.519 3. 738.227. 109. 82 1.m001577 RcCYP74B2 RcCYPN 29901.251. 86.429 274.052 884 1.0.249.227. 191 192.0 1.m014009 28320.m003612 CYP72 clan 30128. 91.868 2.0.m003364 27955. 78.2. 262 0 120.1 0 1. 161.227.0 1.104. 1.429 289. 1. 82 210.0 2.150.364.002. 222.0 1.369.355.242. 95. 207.224.94 215. 1.0.1 2.426 274.2. 147.638 1. 523.251.884 1.917 3.529 1. 225.382.056.426 210.2.RcCYP72A15 RcCYP72A16 RcCYP72A17 RcCYP72A18 RcCYP709B2 RcCYP714A1 RcCYP714A2 RcCYP714A3 RcCYP714A4 RcCYP714A5 RcCYP715A1 RcCYP721A1 RcCYP734A1 RcCYP734A2 RcCYP734A3 RcCYP734A4 RcCYP734A5 RcCYP734A6 RcCYP735A1 29983. 219.0.168. 101 211.m000622 27955.m006256 30170.79. 102. 93. 73.2.1. 109.252.162 1. 100.442 Length of genomic seq’s (bp) Table 1 Organisation of Cytochrome P450 genes in castor 8 8 9 8 0 2 0 4 4 4 4 4 4 4 4 1 9 4 4 4 4 4 4 5 4 4 4 1 No.90.2.379.224.0.1.2.107.251 6.376.035.2. 118.94 194.2.1.236.325.242.340.m000385 30169.376.245.893 3.m000386 29907. 107.2 1.1 1.0 1.m003136 29739. 388.227.2 2.557 274.055 2.899 1.370.

0.914 6 1 3 3 3 2 2 2 2 2 2 3 2 8 8 6 8 7 7 7 7 7 7 8 8 8 No.0. 98.188. 614.132 321.107. 198. 86.249.1. 91.0. 96.0.87.0.448 3.0 2.0 1. 137.m001421 28842. S. 581.79.88 182. 91. 106.108.316 2. 100.188.m000941 29666.442. 178. 211 94.805 2.128. 79.0.1.969 1. 254.1. 263. 384.065 1.456.0 2.128. .m000880 29609. of Intron (s) 135.m001453 29993.1.1.123 RcCYP87A6 RcCYP87A7 RcCYP87A8 RcCYP87A9 RcCYP87A10 RcCYP88A4 RcCYP90B1 RcCYP90C1 RcCYP90D1 RcCYP707A1 RcCYP707A2 RcCYP707A3 RcCYP707A4 RcCYP716A1 RcCYP716A2 RcCYP716A3 RcCYP716A4 RcCYP716A5 RcCYP716B1 RcCYP716C1 RcCYP716C2 RcCYP716D1 RcCYP716E1 RcCYP716F1 RcCYP718A1 RcCYP720A1 30138.128.2 Intron (s) phase 1. 120.2 2.0.297 946. 384. 144. 642.358 3. 124.345 1.300 206.150. 125.0.m000548 30152.0. 99.270. 354 184.0.0 1.93. 67 Intron (s) size (bp) 0. 106.0.2 2.252.0.0 1. 96.649 1.1.0.107.0 1.0. 79.302 2.322. 212.0. 103.325.306 88.0 1.90.1.303 940.328.89.303 451.0. 88 358.0. 213 329.0. 116.0.188.444 5. 98 97. 79.94 197.m000208 CYP gene name 28448. 81 108. 136.114 1.331.m001228 29634. 121. 103.179 501.499 2.322.0. 1.107.150.125 2.m000680 30115.m000795 30074. 79. 101.90. 107. 94.300 928.306 961.0. 79. 84.m003183 28226. 152.234. 297.303 1.0. 107. 107.188.294 943. 96.319.122.322.87.756 3.76. 1. 90 516. 420.107.188.410 2.90.2 2.2 2. 94 233.0.0. 108. 158.97.0.1. 105. 216 106.188. 164. 667.0 1.710 4.343 1.0.478. 84.0.90.992 1. 86 102.671 187 994 1.1.179 931.150.m001052 30172.94 182. 85 402. 163 93.0.0.0.119.733 1.1.0. 107.0.90.m000483 30063.0.0.122.m002401 29728.2 2. 96.309 958.m000359 Castor gene ID Table 1 continued 2. 96.150.1.1.90.0.163 3. 103. 87.0. 82. 81 665 387.188.325.790 2.0.0 1.91 Exon(s) size (bp) 111.119.100 203.m000481 29776. Kumar et al.188.90.0. 538.1. 728.153.0 0.150.90.504.0.119. 482. 117. 79. 186.m002158 29634.109 245. 1. 94.0.273.935 1.227 Length of genomic seq’s (bp) Author's personal copy M.107.m001196 29801. 79. 142 183.m003878 30170.1. 85 100. 88.103 206.m003223 30170.249.84.246. 95.2 2. 113. 86.m002059 28694.0.175.234 233.1.117 665 865 1. 1.566 2.m000875 30018. 107.059 888 807 Sum of intron length (bp) 298 379 478 455 303 473 474 480 483 480 477 480 471 471 455 468 470 482 480 479 492 444 406 479 475 472 No. 102.107.0. 122. 93 127.0.249.0. of amino acids 134 1.0.2 0.0. 154.150.121 1.358.188. 159.372.331.249.508 3. 141.79. 96.1.153.119.014 1.100 200. 87.m013873 29801. 87.0.1.0. 200.0.249.119.0 1. 100 205. 203.0. 159. 283 97.0.128.m000853 29776.387. 241 166. 646 78.153.1 1.0.306 502.893 6.0.790 1.189.0.128. 79.424. 129.m001374 28226.325. 105.m014356 27985.150.2 1 0.249.m000602 29709.0.279.0.97 194.94 200.0 1.204.0.603 2.422 756 1.2 2.021.325.0.918 1.188.234. 145.100 212. 98 96.0.617 3.2 2.115 961.546 391 570 308 268 483 206 183 1.1. 1.150.711 1. 96.2 2.128. 147.779.0. 190.112 221.

120 874 1.0. 158.0.87.0.0.0.111.0.0.252. 33. 295 187. 95.063 Length of genomic seq’s (bp) 5 4 5 4 1 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 9 8 8 8 8 9 No.680 1.84.112 260. 185.0.062 2.524 1.550.m000772 30174.339.201.m002275 29917.0.873 1. 116.267.638 1.216. 143 Intron (s) size (bp) 0.246.m010938 29883. 121.1.119.325. 84.0.107.443 1.319.2 Intron (s) phase 734 1.2.2.0.107.RcCYP722A2 RcCYP722A3 RcCYP722B1 RcCYP724A1 RcCYP724A2 29634.0.153.086.325.1313 1. 2.0. 99. 79.536 3.0.79.180 309. 91.150.m001310 CYP86 clan RcCYP722A1 CYP gene name 30170.m000931 30005.0.569 2. 87.563 1. 244.2 2.921 1.m004790 30147.555 2. 51.252.0 0.0 2.0.409 110 0 0 1. 87.1. 79.m000268 29660.1.90.m014517 29883. 628. 79.0.0.87.867 4.518 1.1. 979 149.m000137 29811.0. of amino acids Author's personal copy Organization and Classification of Cytochrome P450 135 123 .0. 1.521 1.0.0.107.530 1. 89. 79.285.067 4. 562.0.0 0 0 1 2.500 1.100 146.107.393 256.0.m001270 RcCYP86A3 RcCYP86B1 RcCYP86C1 RcCYP94B1 RcCYP94B2 RcCYP94B3 RcCYP94B4 RcCYP94C1 RcCYP94C2 RcCYP94D1 RcCYP94D2 RcCYP96A1 RcCYP96A2 RcCYP96B1 RcCYP96B2 RcCYP704A2 RcCYP704A3 RcCYP704A4 30174.m002092 29863. 213.498 511 507 203 0 0 0 0 0 0 0 0 0 0 0 0 1.536 221.m000531 30078. 89.m002010 29409.m001518 RcCYP86A1 29681.1.153.305 2.521 1563 1506 1.530 1.168 522. 455.0 1.m000683 29929.119.325.613 1.m000057 RcCYP704B1 RcCYP86A2 30190.m001089 29982.156 200.107.1. 349 111.122.832 2.530 1. 94.2 2. 226 257.322.26.150.119.494 1.391 2.046 1.119. 288.m008915 30194.588 2.0.0.563 3. 206.0.91 248.m002015 30190.443 1.0.243. 201.0. 219.0.80.100 Exon(s) size (bp) 179.409 110 0 0 129. 86. 95 203 0 0 0 0 0 0 0 0 0 0 0 0 1. 98.144.136.150.494 1. 90.m000529 28779.216. 84 85. 107.m002017 29791.563 1260.686 Sum of intron length (bp) 536 394 347 512 469 505 505 509 506 520 501 499 507 509 497 480 520 550 522 545 511 523 482 476 491 492 458 No.m008914 30174. 321.m008617 30094. 104.m011234 29813.0.404 3. 95. 97 217.045 1.518 1.297.291.0.192 1.345 2.m014078 Castor gene ID Table 1 continued 2.m002008 29917.2 2. 387. 831. 111. 101. 289. 256.76.m000224 29633. 227. 105.0.0. 87. 109. 337. 129.324 1. 93. 106.178. 88.92.153. 99.45 277. 89.184.1.530 1. 388.2.0 0. 79.0. 76.87.518 1.0. of Intron (s) 330. 133 325.0.1.506 1.0. 723 108.325.500 1. 183.115 209.518 1.

153.445 4.0.123 RcCYP97C1 30078.102. of Intron (s) 891. 81. 549.m002224 28223.66 71.0. 2.0 445.0.0.2.0.621 945. 213.132. 587.83.0. 593.621 477.2.720 2.m013848 30129.290.0. 328.0 0 0.865 1.151.0.m013846 CYP71 clan 29686. 202 1.475 2. 141. 192.m006275 RcCYP727A1 RcCYP711A1 30147.0.87.83.85.146 Length of genomic seq’s (bp) Author's personal copy M.621 915.0.150.m013843 30147. 154.1.627 882.0.670 17.m000959 29785. 0.m006288 29785.0.127. 713.886 3.501 2. 97 88 477 975 338 187 918 255. 111.1.347 1.163.89.560 1. 168 Intron (s) size (bp) 102 95 132 251 493 268 112 129 79 892 856 690 138 214 88 477 975 338 187 918 3.0.627 123.m000100 RcCYP71A24 RcCYP71A25 RcCYP71A26 RcCYP71A27 RcCYP71A28 RcCYP71B1 RcCYP71B4 RcCYP71B8 RcCYP71B9 RcCYP71B10 RcCYP71B11 RcCYP71B13 RcCYP71B14 RcCYP71B22 RcCYP71B23 RcCYP71B24 RcCYP71B25 RcCYP71B26 RcCYP71B27 RcCYP71B28 30147.662 1.217. 400.618 915. 107. 192. 1.108.621 882. 581.559.0 2. 1.72.609 945.623.615 501.33.0.0. 96.m009010 CYP97 clan Castor gene ID Table 1 continued 1.624 666.66.612 208.136.621 996.329 19. 255 268 112 129 79 892 856 79.638.199. 99 117.608 1.78 248.108.1. .734 1. 1.466.1 0 0 0 0 0 0 1. 174. 100.200.m000821 30169.153.615 1. 0.m000757 30169.647 1. 276.247 Sum of intron length (bp) 501 506 504 500 438 507 500 534 532 509 509 480 473 315 518 527 521 508 510 508 512 536 159 552 555 632 No.60.624 906.615 894.784.2. 148.678 2.111.48.330 903. 233.m000239 29910.0.359 942.541 1.612 915.m000867 CYP727 clan 29739.178.061 2.26. 128.616 1.0.0.m013842 30147.m000240 29887.0 Intron (s) phase 81. 0 0 0 0 0 0 0 0.101. 328 630.m000965 29785. 77 143 0 0 0 0 0.m013847 30147.0 1.m000239 29887.102.2.m000853 CYP710 clan RcCYP97A3 CYP gene name 30128.615 906.m003566 CYP711 clan RcCYP710A1 RcCYP97B3 29724.645 2.0 0. 390.m000241 30169. 137.297.0 2.m000355 29724.239.0.189 Exon(s) size (bp) 102 95 132 251 101.m000962 29826.665.198. 92. 1.469 5. 68.225 139.96. Kumar et al.754 1.0. 64.0.0. 611 39. of amino acids 136 1. 486. 105 559.615 963.185 1. 592.62.2. 89.m000943 29878.m006277 29887.0. 116.642.590 143 2.615 897.810 1 1 1 1 3 1 1 1 1 1 1 2 2 2 1 1 1 1 1 1 6 4 1 8 13 15 No.162. 165. 681. 10.624 157.2.029.2.61. 671. 200. 462.m006273 30169.0.792 1.201 480 4. 148. S.163.654 906.801 3.

615 Exon(s) size (bp) 0 0 128 153 149 107 93.037 176 97. 605 906.1 0 0 0 0 1 0 0 0 0.633 1.m000520 29428.609 903.725 1. 736 785.615 930.m003625 CYP gene name 30169.0 2 2.m014296 30169.596 930.m000966 29929.m000509 29815.51 885.245 942.682 1.313 1.615 1.624 900.815 981 2.616 1.944 2.013 2.065 2.673 1.m004562 29792.630 927.21 906.612 862.794 1.722 2.m000516 29815.1 0 0.m004748 29976.445.339 1.m004561 29629.701 723 1.210 2.m000643 29792.651 879.612 885.729 Length of genomic seq’s (bp) Author's personal copy 137 123 . 599 927.m001392 29929.m003983 30146.m000783 29785.m003563 29706.m011069 30147.624 906.996 1.472 250. 1.632 1.m006279 Castor gene ID Table 1 continued 1.0 0 0 0 0 0 1.630 942.399 2.679 1.472 302 824 150 108 132 269 208 93 122 157 Sum of intron length (bp) 505 531 514 515 509 524 512 502 514 525 501 426 318 496 496 501 377 521 511 521 461 536 505 520 499 304 507 362 504 524 507 499 504 512 533 497 523 No.596 1.037 176 210 499 142 121 84 179 156 547 573 1.m006285 29792. 510.682 1.035 66 1.12 906.695 1.m006295 30169.m006291 30190.m011130 29815.973 3.612 1.m000519 29815.0 0 0 0 0 1. 87 903.m000625 29929.651 875. 285.139 3.744 362 1.63.609 900.491 1.m000510 29815.456 1. 312 556 468 1. 52 824 150 108 132 269 124.636 903.m000508 29815.130 1.618 939.m003754 30190. 853 362 176.609 915.518 1.518 0 0 1 1 1 1 2 1 1 1 1 2 1 1 1 1 1 1 1 1 2 1 2 1 2 1 1 2 1 1 1 1 1 2 1 1 1 No.747 1.m001350 29629.624 891.632 1.0 0 0 0.576 2.315 66 1.m006290 30169.624 282.m000626 30142.391 2.m000624 30206.627 391.609 957.1 0 0 0 Intron (s) phase 0 0 128 153 149 107 405 556 468 1.m001271 29739.614 990.525 723 280.606 882.621 900.591 960. of amino acids Organization and Classification of Cytochrome P450 1. of Intron (s) 1.m000515 29815.606 889. 113 499 142 121 84 179 156 547 573 891.615 473.701 1.m011068 30190.453.174.m000318 29842.618 894.609 951.784 1.m006282 30169.m000504 43540. 84 93 122 157 Intron (s) size (bp) 0 0 0 0 2.RcCYP71B29 RcCYP71B30 RcCYP71B31 RcCYP71B36 RcCYP71B37 RcCYP71B38 RcCYP71B39 RcCYP71B40 RcCYP71B41 RcCYP71B42 RcCYP71C1 RcCYP71C2 RcCYP71C3 RcCYP71D1 RcCYP73A5 RcCYP73A6 RcCYP75B1 RcCYP75B2 RcCYP75B3 RcCYP75B4 RcCYP76C3 RcCYP76C4 RcCYP76D1 RcCYP76D2 RcCYP76D3 RcCYP76D4 RcCYP76E1 RcCYP76G1 RcCYP76G2 RcCYP76H1 RcCYP76H2 RcCYP76I1 RcCYP76I2 RcCYP76I3 RcCYP76I4 RcCYP77A4 RcCYP77B1 30169.m000048 30138. 1. 134.m000512 29815.

1.m003950 29929.621 903.m013958 30170. of amino acids 138 1.630 945.113 1.684 0 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 0 1 1 1 1 1 1 1 1 2 0 0 1 0 1 1 1 1 1 1 0 No.m001679 30170.m013953 30120.m013949 30170.m000134 28644.624 948.166 533 349 94 92 157 0 Intron (s) size (bp) 0 0 0 0 0 0 0 0 0 0 0 0 0 1.627 945.529 Length of genomic seq’s (bp) Author's personal copy M.166 533 349 94 92 157 0 Sum of intron length (bp) 313 441 496 497 523 528 522 521 523 526 521 521 530 525 495 528 320 505 319 504 500 509 503 514 517 452 338 370 329 390 519 462 531 537 533 340 505 No.624 939.636 996.464.017 1.621 927.m002578 30138.518 Exon(s) size (bp) 0 1.m000371 30170.223 3.071 1.936 2.618 1.045 1.612 885. .673 2.180 1.790 13.621 948.m007121 CYP gene name 29842.0 0 0 0 0 0 0 0 Intron (s) phase 0 1.m013965 30120.9 1.584 2.m001002 30170.m013950 30170.611 2.726 2.612 879.708 1.699 963 1.123 RcCYP77B2 RcCYP78A6 RcCYP78A7 RcCYP78A8 RcCYP78A9 RcCYP78A10 RcCYP78A11 RcCYP79A2 RcCYP79B3 RcCYP79B4 RcCYP79B7 RcCYP81D2 RcCYP81D3 RcCYP81D4 RcCYP81D8 RcCYP81D9 RcCYP81D10 RcCYP81D11 RcCYP81H1 RcCYP81K1 RcCYP82C2 RcCYP82C3 RcCYP82C4 RcCYP82C5 RcCYP82C6 RcCYP82C7 RcCYP82C8 RcCYP82C9 RcCYP82C10 RcCYP82C11 RcCYP82C12 RcCYP82C13 RcCYP82G1 RcCYP83B1 RcCYP83B3 RcCYP83B4 RcCYP84A1 28014.m014153 30170.633 903.005 306 117 1.026 3.217 158 624 406 1. 207 112 0 187 12.627 891.609 942.253 1.m000917 29910.797 1.072 766 1.m013960 30170.m014207 29851. of Intron (s) 942 714.618 999.146 1.m013773 30170.694 1.072 766 1.287 2.597 783.173 963.005 306 117 1.848 2.636 272.311 3.m013957 30170.m000914 28438.331 1.694 1.472 1.469.m014208 29970.017 1.542 122 267 721 580 53 1.m000050 29970.621 327.m014151 30174.603 405.m002485 30170. S.621 930.m009168 30131.m000998 30170.m000933 30068.217 158 624 406 1.630 963.624 954.112 145 654 1.633 963 897.627 400.158 297 112 0 187 12.m004656 29910.m000118 28256.127 1.0 0 0 0 0 0 0 0 0 0 2.745 0 0 81 0 1.666 951.112 145 654 1.472 149.m000372 30170.732 2.m013780 29970.606 960.950 1.m013964 30170.m000911 29910.633 939.m000948 30170.108 1.177 1.646 2.618 1.736 1.113 981.m001003 29910.173 2.785 1.542 122 267 721 580 53 1.158 90.894 0 0 81 0 1.612 882.615 918.m003626 Castor gene ID Table 1 continued 942 2. Kumar et al.m013963 29676.m013774 30170.

m000401 30147.m000400 30170.783 2.m000323 29940.m001476 30148. 314 178. 84. 1.434 2.m011007 30147.m014117 29940. 163.2.542 1.959.545 1.093 143 Sum of intron length (bp) 516 518 513 491 518 530 515 513 395 511 511 506 333 546 409 493 390 514 500 518 514 513 524 520 516 516 501 511 505 No.568 1.0 0.m001475 30148.574. 83 2.575 1.671 1.909 709 294 1.536 493. 335 70.204 898 0 0 0 0 0 0 0 0 0 0 277 1.m000255 30152.173 1.093 143 Intron (s) size (bp) 0 0 0 0.503 1.153 1.398.551 1.347 936.0 1.2.630 939. 824.551 1.116 112 340 185.m000256 29216.m014189 29216.230 1.988 4.0 0 1.822 903.m000174 29216.m000257 CYP gene name 28196.694 3.116 112 340 1.428 811.681 891.2.666 802.m001482 30148.151 3.542 1.m001483 29083.633 909.m001406 30190.235. 158. 85.m011008 30190.738 1.820 1.551 1.m002236 30148.551 675.m000321 29788.m001478 30148.563 1.311. 973 0 1.645 609.m001481 30073.204 898 0 0 0 0 0 0 0 0 0 0 277 1.380 1.111 16.281 425 172. 102.621 960. 639 294 157.630 894.m001477 30148.639 154.456 3.536 3.1 0 0 0 1.233 1.557 1.705 1.618 912.503 1.615 Exon(s) size (bp) 600 1.472 1. 160.612 1.551 1.031 2.398.583 2.140.m003926 30174.661 Length of genomic seq’s (bp) 1 1 1 2 1 1 1 7 2 0 2 2 1 2 1 1 0 0 0 0 0 0 0 0 0 0 1 1 1 No.0 1 1 0 0 0 Intron (s) phase 600 1.1.296 3.m008711 29827.0.m000205 Castor gene ID Table 1 continued 2.390 1. of amino acids Author's personal copy Organization and Classification of Cytochrome P450 139 123 .m000045 29216.888 2.828 714.1 1.281 425 218 2.284 0 1. 258.m002605 30148.m000258 27647. 93.601 1.545 1.545 1. 195.231. of Intron (s) 921.967 1.m013942 29742.575 1. 46 2.RcCYP84A2 RcCYP84A3 RcCYP84A4 RcCYP89A4 RcCYP89A5 RcCYP89A6 RcCYP89A7 RcCYP89A8 RcCYP89A9 RcCYP89A10 RcCYP89A11 RcCYP89A12 RcCYP89B1 RcCYP93D1 RcCYP93D2 RcCYP93D3 RcCYP93D4 RcCYP93D5 RcCYP98A3 RcCYP98A4 RcCYP98A5 RcCYP701A3 RcCYP703A2 RcCYP706A4 RcCYP706A5 RcCYP706A7 RcCYP712A1 RcCYP712A2 RcCYP712B1 30138.m002423 29788.629 1.

m 3563 C At YP 0037 C 5 7 A 4 0 Y At A tCY P7 5A1 CY tC P7 05A 6 Y 0 P P 5 19 A 705 705 A23 At At tCY A21 A20 CY CY P7 A P P7 05 tC Y At 705 05A A15 P7 C A 05 YP 8 4 A 70 5 5A 9 Fig.m 74 YP 2A1 12B1 001 -297 AtC YP71 CYP7 712A2 647.m0 0129 842 B 3 3 7 3 3 P8 83B 4-301 71B1 7.m00 000240 B YP7 1B31 14-30169 0241 -3 . CYP703.m RcC 169 0190 C7 2-30 4-3 P76 3 76D P76C 8 CY P76C 295 t P 6 Y 0 A Y 6 0 Y C6 RcC RcC 0110 9.m 01 9 4 0 Rc c 0 R P8 301 . CYP74.m0 70.m0001 33 34 RcCYP78A1 AtCYP78A7 0-30138.m001 0100 YP 00 2 AtC 81F4 3 YP A RcC AtC tCYP8 81F3 YP8 YP8 1F2 1D RcC 4-30170 AtCYP 1F1 Y .m cC 00 cC 71 9-29 -29 785 R 5. A4.m00 -2 P71B 0239 26-2 9887.m P 0 26 Rc 77B At 003 0004 05 CY 2 C P7 -298 YP 625 5 4 7A 7 4-2 2. and A2.m0 13847 13848 1 Rc P7 A26 147 0 0 7.m 000 YP 096 Y C 78 78 . 914 1 7 000 9 000 10.m006 . At tCY YP 81D m At CY P8 81D 3 At 00 CY 3 CY P8 1D 2 1D 8 P P7 983 At AtC AtC 81D 10 5B At CY YP YP 5 P 1 Rc C P 8 8 YP 81 1D 1D C Rc 81 D7 11 4 CY YP8 D 1 P8 1D H1-2 AtCY AtC 6 3-2 997 P8 YP Rc 9 C 1 RcC YP 910 0.m0111 006291 RcCY E1-3019 0169.m 000 2 -29 -29 AtC 71 88. A8.m00 76 P 30 RcCY P76G1-290.m Rc cCYP 1A24 -3014 8-3014 7 7 2 R YP A2 1 71A C 7 P c P Y R CY 1 RcCP71A2 2 Rc 2 Y AtC YP71A 4 AtC P71A2 Y 6 AtC P71A2 Y AtC P71A23 AtCY P71A25 AtCY 71A12 P AtCY YP71A13 AtC 1A18 AtCYP7 1A19 AtCYP7 20 1A AtCYP7 YP71A16 AtC 27 AtCYP71A AtCYP71A28 AtCYP71A14 AtCYP71A15 RcCYP71D1-29929. C1.m P7 00 1B22 6279 RcC RcCY YP71B25 -29887.m 0 0 AtC . 940.The families CYP83.m00 AtCYP82F 1679 1 RcCYP82C5 RcCYP82C2-298 -30120.m 8 RcCY P81D11-3 014208 1G1 P81D 0170 9-30 . D1. orthologs A1.m RcCYP7 6H2-29 10 RcCYP7 815. and CYP735 represent conserved number of genes between the genomes of castor and Arabidopsis.m -299 9910.m 1008 706 A1 07 7 14 01 YP 06 0 -30 .m R P 0 8 9 1 B 1 97 A CY 71 1B 1-2 3-2 83 Rc YP P7 1B1 3 1C YP 15 C Y 7 P7 AtC 3B1 014 4151 5 Rc RcC CYP 8 . B1. A7. RcCY m0 RcCYP7 P71B40-29929.m 09 095 21B cC 79 00 00 962 71C 6 P7 R1-29 5. CYP721.m 00 00 9A 301 014 8. A6.148 9 R 9A -30 P8 P8 A10 CY 89 Rc YP cC R Rc A AtC At tCY YP Rc At CY P8 89 P 9 A Rc CY C P8 At YP 89A A2 3 C Rc CY YP8 9A4 At At CYP 89A 7 9 P7 7B B1-2 2982 CYP CYP 89A 6 1 9 8 7 8 0 .m00 Rc Rc C At 1-3 3017 4.m P 42 Y AtC 2 97 AtC 9F 2-2 P7 F1 3A 1 CY 79 At CYP 79C2 79C P70 At YP CYP cCY C R At At 0.Y Y Y 06A 5 Rc P70 6A AtC AtC AtC P7 6A 70 CY 0 CY 7 Rc YP C At YP Rc tC A R 2 2 A A3 28 05 05 5A 30 P7 P7 0 5A Y Y P7 70 2 tC tC Y P A2 A A AtC tCY 705 3 A P 5A 0 CY P7 A6 8 At CY 705 5A1 33 At CYP P70 05A 3 At tCY YP7 5A1 2 24 A AtC P70 A1 A1 5 5 705A CY 70 70 At CYP YP CYP At AtC At 25 05A 258 00257 P7 A27 000 0 CY 05 321 At YP7 A1 216.m 74 01 0.m tC P7 011 A7 0190 A tCY 0.m0 00 14 22 9-3 48 8. and nine paralogs (A3.m 1 8 CY 035 YP 0170 0.m 15 RcCYP76I2 6I1-29815.m 00 06 25 Rc C 76 14 00 78 7 .m 79 B 06 20 29 P71 00 65 9 0 3 41 Y 2.m004656 RcCYP78A9-30068. CYP98. CYP718.m000943 878. 9 Rc 29 9 5 84 83.m 216. and CYP84 exhibited two additional genes in castor as compared to the Arabidopsis.m 71 11 1.m0005 6G1 29 tCYP7 09 P76H1A Y cC R 0005 08 05 815.m 74.m 00 84A P7 -30 00 00 3926 1-3 5B 14 12 87 013 4-2 6.m000757 RcCYP71B38-29629. C2. CYP704.m011 . and A10) were recorded in the castor genome.m CY 3.m0 00371 AtCYP82G1 RcCYP82C13-30170.m 5. ‘‘Rc and At’’ represent CYP proteins of castor and Arabidopsis.m0004 m 00 00 RcCYP7 AtCYP90401 8A8-286 8A3 RcCYP7 44.m01 3843 Y P B 01 4 C P Rc cCY P83 cCY 5-301 147.m01 3780 37 P81K 1-3017 -30170.m 004562 006273 1B RcCY 29-30169.m0 006277 P 0 AtCYP71B 71B28 P71B4-2 6285 YP 30-3 -301 9724 84A 6 016 Rc 1 9.A4 301 00 RcC Rc P75 297 -301 38.m000239 RcCYP71B RcCYP71B 8-30169. CYP76.m CY 2-3 Rc A1 cCY 11. Arabidopsis CYP716 family showed two genes (A1 and A2) while castor displayed 11 genes viz. and CYP712.m013949 9-3 RcCYP82C m013950 10-30170.m 9.m0 7B1 9 0 AtC 428.m YP7 000 3A6 A 5 -435 40. A5.m 3846 1 R CY R A2 -30 .m0141 RcCY YP98A5.m0005 000512 815.17 P98A329 29940.m m00 A9 A4 CY 2.m 04561 1B RcCYP7 27-30169.m 0 RcC 2-2991 438. S.m AtC P76 C5 6C1 016 . 42 24 6.m01 R RcCY cCYP81D10 170. E1 and F1). CYP715.m 7 013 tCYP A6 77A AtC 942 4 Y RcC P701A YP7 3 3A5 -299 RcC 76.Author's personal copy 140 M.m013963 RcCYP82C11-30170. AtCYP8 m014207 3773 1K2 AtCY RcCYP8 2C8-296 P81K1 76.m000118 AtCYP78A8 AtCYP78A9 AtCYP78A6 29815.m0 0916 . 216. CYP707.m 0205 YP CY B2 06.m 02423 -297 0 3D5 0152.m001350 RcCYP71B39-29629.m 96 -3 4 76 6D 0142 YP 7.m CY 71 -30 7. Kumar et al.m 3 P8 8-3 14 48 3.m0 6-30 00 482 C CY P89 148 A 8. CYP51.m000520 6I4RcCYP7 5.m R RcC cCYP71 9887. .m A7 A3 A4 6 0 A 19 706 706 706 6 P7 4-3 30 P P P CY 6A 5.196.m YP9 -3 56 RcC P93D3 D1 m0002 Y 93 0255 RcC AtCYP D1-29216.m 75 CY 9A A7 -30 01 07 14 Rc YP8 P89 A5 .m 9B 4 -2 911 YP7 9B3 000 00050 RcC YP7 0.m000 51.m00 09 8A7-282 56.m006288 37-29929.m001392 RcCYP71B RcCYP71B24-29 23-29910.m013965 RcCYP82C12-30170. CYP97.m013953 2C4 2C4-3 AtCYP83 RcCYP8 2C AtCYP8 2C2 P8 AtCY P98A 3 64 00 6 m0 62 42.m002578 RcCYP78A6-28014.m002485 372 RcCYP82G1-30120.m013958 RcCYP82C3-301 0170. A4.m013964 RcCYP82C7-30170.m 362 RcC YP 00 6 YP 701 AtC AtC 77A9 0318 A3-3 YP YP7 017 7A 77 A 0.m013960 13957 RcCYP82C6-30170.m H 81 D1 YP 81D .m YP7 RcC RcC 1-3014 6D 069 290 YP7 006 . respectively Comparative analysis of CYPs revealed that the families.m00082 C 2 006 AtC 1 282 75 R YP Rc Rc cCY 84A2 YP84 CY CY P8 -28 A4 Rc P75B P84 4A3. As compared to Arabidopsis. 1 Phylogenetic tree of A-type Cytochrome P450 proteins of castor and Arabidopsis.1 AtCYP71B21 AtCYP71B22 AtCYP71B38P AtCYP71B5 AtCYP71B30P AtCYP71B AtCYP71B 32 31 AtCYP7 AtCYP71B8 1B23 AtCYP7 1B7 AtCY AtCYP7 1B15 P71B AtCY AtCYP71 28 P71 B29 AtC B2 YP7 1 A AtC tCYP7 B27 AtC YP71B 1B16 17 Y AtCAtCYP P71B1 9 Y 7 AtC P71B 1B20 AtC AtC YP71 24 YP YP7 B3 71 1B A B4 25 A tCY A t tCYP P 7 1 B CY 71 11 A P B A t 71 12 A tC CY B1 A tC YP P71 3 At tCY YP7 71B B14 At CY P71 1B2 6 B 6 C P A Y 71 33 At At tCY P71 B34 B At CY CY P71 35 P P R CY 71B 71B B9 cC P7 3 3 Y 1B 7 6 P7 1 1B 0 36 -2 97 92 . 123 Similarly CYP93 family showed five genes against one in Arabidopsis.m 79A B7-28 YP 79 YP RcC CY 2 79A YP 3 AtC P79B 2 06 2 14 A Y 9B 00 703 AtC YP7 . The family CYP87 consist of 1 gene (A2) in Arabidopsis whereas 8 genes (A2.m000519 981 3-2 RcCYP76I 000516 -29815.m 014 147 1 8 48 36 01 .m004748 RcCYP71B13-29826. RcCYP82C 0170.m 97 00 007 121 At 39.. About twofold amplification of genes is observed in the families.m Y 76 P7 190 D3-3 C t 6 0 7 P A Y tCY 6C4 6C2 3-3 YP C A P7 7 76C RcC At P YP CY Y RcC At AtC 89 41 A2 01 . 0 83 00 01 07 m -3 00 2. castor genome exhibited an additional gene in the families CYP73.m RcC RcCY RcCY 0169.m tCYP7 04 0000 3A5 48 RcCY AtC Y 4-3014 AtCYP P98A9 98A8 RcC 7.m0 000 1 81D 8-2 00 9 2-2 997 948 98 997 0. A9. CYP701.m0 03950 AtCYP78A10 AtCYP78A5 RcCYP78A11-29929.m0 P93D4 P RcC Rc AtCY 2A2-27 RcCY 1 YP7 RcC 3 2 0003 88. CYP78.m00 P93 D2-29 RcCYRcCYP93 cC Y P7 5B At 130 C A 13 A tC YP 8. A5.m G2-29 815.

m 94 00 -3 B1 790 004 1 9. CYP706.m 4B2 -299 YP7 4B2 AtC YP 7 RcC 972 013 74A 0.m 7 8 985 000 .m CY P86 2 P7 A4 00 04 P8 89 A3 6A 1 4 30 8 17 4.m F1 63 8A 42 300 16 71 8 7 P 8 P 1 -2 CY D1 048 CY E1 Rc 16 Rc 00 16 P7 .m 8.Author's personal copy CY P8 P8 Rc 0.m AtC C4 -296 P86 6B2 Y 08 YP9 0020 AtC .m 1-2 .m 707 1 P 70 0 Y 98 YP 1 2 C RcC t 7A A A2 70 707 YP YP C t C A Rc 18 P7 3 52 CY 145 10 At 00 00 21 m .m CY 2A 2 R cC At 70 309 A P 02 PN CY P7 CY At A1 Rc CY 02 8 At P7 2A CY P7 0 3 At A1 8 A CY 70 02 At P7 YP A4 CY AtC 08 At P7 3 CY 8A 0 At P7 Y tC A Rc Rc 141 AtCYP94B3 Organization and Classification of Cytochrome P450 10 AtCYP90A 1 -29634.m001089 RcCYP722A3-29863 092 29634.m0 734 4 6 6 YP 017 090 RcC A2-3 . The observation is in conformity with CYPs representing 0. 7 01 3 CY 7A 7A 7A Rc P8 0. CYP90.m00 2A18-29 RcCYP7 1 AtCYP72C 12 4-29739. 19 0 -3 P8 CY Rc 00 01 01 P8 CY Rc R 8.m AtCYP72A7 AtCYP72A8 AtCYP72A9 AtCYP72A 14 AtCYP7 2A10 AtCY P7 2A 15 AtCY P72A11 AtCY P72A 13 RcCYP72A1 95 P CY 716A P7 2 2. poplar 123 .m 22B1-2 P7 Y RcC 88A4 P Y AtC 3 P88A AtCY 1228 83 .m 72 07 Rc P7 03 0 .m0 7A2 5 1 1 70 223 -30 YP A1 003 7A3 AtC .m RcCY AtC P90D 0002 YP7 1-2869 08 20A1 4.m 77 69 15 01 00 -3 . CYP86.m 2A16-2 3138 RcCYP7 983.m00 AtC 9010 YP9 7B3 RcC YP9 7B3 -297 24.m 34 12 01 2 Y cC A2 44 14 19 CY 08 28 30 30 3- A 6 P8 P7 6- 5- 2- 17 86 00 m 4.m 79 99 4 4 00 Y 4 .m 1 RcC 000 YP7 806 20A 1-30 172.m 7A P8 7A 10 8-3 -29 01 CY P8 70 9.m002 RcCYP722A20.m A1 5A1 A2 05 3 15 5 02 -3 P7 YP7 P73 A1 CY tC tCY 35 B1 A Rc A P7 09 CY YP7 534 B2 Rc C 004 09 67 0 At P7 3 6.57–1.m002 158 AtCYP96A RcCYP90B1 AtCYP96A9 AtCYP90B1 AtCYP96A1 RcCYP724A2-30005.m000931 AtCYP96A7 AtCYP724A1 AtCYP96A8 AtCYP96A13 AtCYP96A 12 AtCYP9 AtCY P97A 3 P97A 3-3012 8.m -2 55 A 4 9 00 -2 tC .m0 174 734 YP 3-30 34A RcC 1 YP7 21A 9 0 RcC YP7 0140 AtC 70.m At 1-2 000 CY 973 100 P7 9.m0 P73 174 CY 65 -30 Rc A5 090 .m CY P7 001 At 04 P7 518 CY A1 04 A4 P 70 Rc -30 4A CY 19 2 4.m00 0680 AtCY RcCY P90D P90C11 29634.m0 RcC 07A 7A4 170 YP7 P70 3-30 Y A RcC 7 6 AtC P70 119 Y 0 C Rc . However phylogenetic tree displayed distinct projection of this gene from CYP74 family (Fig.m 00 01 2 06 43 02 56 38 . respectively CYP82 and CYP89 of castor as compared to Arabidopsis. A single intron containing non-A-type castor gene RcCYPN coding for 416 amino acids (Table 1) exhibited 42. 96B1P Y RcC 3 P96A AtCY 6A4 AtCYP9 6A15 AtCYP9 6 P8 CY 6C P8 CY At AtCYP9 B2 86 P CY At 83 06 1 6B P8 Rc 798 .000).07 % of the protein coding genes of Arabidopsis (246/23.m 000 008 875 568 AtC YP 51G AtC 1 YP AtC 51G YP7 2 10A AtC 4 YP7 10A 3 AtC YP7 10A 2 AtCY P710A 1 74 13 00 00 .m000932 Rc 6A11 AtCYP9 6A2 56 62 00 .m00 9 0 7 0031 -29 73 801.m 00 03 85 53 07 00 A3 8. Discussion The present study on the genome analysis of castor revealed the presence of 210 putative CYP genes belonging to 45 families.m -30 16 A2 16 16A P7 P7 CY CY Rc Rc Rc C YP 5 P7 CY Rc 01 -30 15 At At CY . and CYP710 of castor exhibited less number of genes as compared to Arabidopsis.m 74 00 -3 A5 16 16 P7 CY Rc CY R P7 cC Y B1 P7 -2 16 82 C 26 2- 29 . 2).m0036 .m 77 6.m 7. m0020 59 Rc Rc 6A P CY At m 0. CYP81.m RcC 7 1 -299 6A1 10 YP9 0020 RcC 917.m00 AtCY 1-2832 P734A 003136 RcCY 9983. rice (356/37.5 % similarity and 30.68 % of the predicted *31.m 00 38 5.m P7 CY 776 CY 9 Rc Rc 2 1C 16 P7 Y cC R Fig.m 81 96 -2 A1 CY At 10 13 00 A1 86 YP C At CY 7-3 7A A4 A7 86 Rc P8 7A .m 42 11 At 60 014517 RcCYP94B3 -30190.m AtC 0008 YP9 5 7C1 RcC YP 97C 1-3 007 8. CYP72.m At Rc 003 CY 14A1 CY 566 P7 P Rc Rc 727 14 CY CY A2 A 1-2 Rc P7 P7 Rc 968 14 CY 14 A1 6.m 00 89 15 Rc At CY CY 415 000 01. 08 m 00 04 83 RcCY RcCYP72A15-29633. 6 41 014 3 66 99 09 0 9 9 0 0 -2 . CYP96.m00 RcC 05 29 YP9 4C2 RcC -287 YP9 79.m014078 22A1-3017 RcCYP7 22A1 AtCYP7 000224 9982.m010 938 AtCYP94B 2 AtCYP94B1 RcCYP94B1-30147.m A2-29 68 YP96 m0002 RcC 29409.m 16A 1 00 24 01 1-2 1G 00 1-3 822 05 012 48 6. CYP77.544) [14].m -301 21A1 YP7 RcC 1 78 10 P734A 0.m 992 1-2 6C 8 P 2 CY 86C Rc YP 3 AtC 772 86C 000 YP 60.m001270 AtCYP96A5 RcCYP724A1-29633. 2 Phylogenetic tree of non-A-type Cytochrome P450 proteins of castor and Arabidopsis.m 4D2 AtC 0005 -3 007 YP 31 8. grouped into ten clans. m 0 86 CY 15 33 00 A 64 06 P7 1 22 14 A5 -2 79 55 .m At 2-3 P 068 174 009 -30 CY 09B A4 At P7 4.m YP 017 6 A tC 018 1-3 02 A 000 74A P7 5 YP 10.m P7 A3 CYP -29 000 14 -2 71 84 867 A2 79 4A 8. m .m 03 59 92 35 CY 7A CY CY P8 P87 -27 01 9-2 6 P8 0C1 00 . The gene families CYP71.m P7 At 00 04 00 CY A2 At 57 -30 P8 CY Rc 6A At 17 CY 4. ‘‘Rc and At’’ represent CYP proteins of castor and Arabidopsis.m 4D1 RcC 0001 -298 YP9 37 11.m0 880 008 78 AtC YP RcC 85A YP8 AtC 2 5A1 YP -297 85A 90. CYP450 genes identified in castor represent *0.m 4 3 017 CY P7 70 6-3 Rc 090 CY 4A Rc .8 % identity with RcCYP74A1 gene.m 009 CY 709B 007 .m 8A4 138 YP8 4-29 .m AtC RcC 002 YP 224 YP 711 Rc 710 A1 CY A1-2 P7 822 11A 3. m0020 AtCY 17 P94C 1 RcCY P94C 1-2979 1.2 RcCYP9 4B2-298 83. CYP79.m00 2015 RcCY P94B429883.m 94D AtC 002 1 275 YP 94D A tC 2 YP 704 Rc B1 CY P7 04B 1-2 At 981 CY Rc 3.m 28 97 -2 A4 16 P7 CY Rc 003779 RcCYP72A17-29739. CYP709.000 protein coding genes.

) a review. Cahoon EB. S. Schuler MA. Beisson F (2011) Cytochrome P450 metabolizing fatty acids in plants: characterization and physiological roles. Wortman JR. Redman J. thereby limiting their diversification. Mahasi MJ (2008) Role of biotechnological interventions in the improvement of castor (Ricinus communis L. Sujatha M. Stecher G. pp 13–52 2. Oxford. Franck P (2011) Cytochrome P450 metabolizing fatty acids in living organisms. These results are in conformity with the earlier report indicating the absence of whole genome duplication in castor. Lorenzi H. 710. As compared to Arabidopsis. Biores Technol 97:1086–1091 5. Anzenbacher P. Werck-Reichhart D (2011) A P450 centric view of plant evolution. Crabtree J. Chen G. Single-family clans CYP51. Tamura K. Pinot F. Chan AP. Ogunniyi DS (2006) Castor oil: a vital industrial raw material.Author's personal copy 142 (310/45. Zhao Q. Papi Reddy former Head. Edgar RC (2004) MUSCLE: mutiple sequence alignment with high accuracy and high throughput. 711 and 727 represented with a few (1-3) genes are plausibly ancient and may code for enzymes associated with essential functions. Haas BJ.434). Guerci A (1982) Various uses of the castor oil plant (Ricinus communis L. 97. Nelson DR. The gene might have diversified from RcCYP74A1 gene. soybean (332/46. Occurrence of more number of A-type (123) CYP genes as compared to non-A-type (87) in castor implicates to the rapid expansion of A-type CYP genes over non-A-type. indicate it as a novel gene. rather than to changes in the whole chromosome complement [19]. Puiu D. cryptic inversions. Blackwell Science. The highest number (123) of the CYP genes are A-type and the remaining 87 genes are non-A 123 M. papaya (142/24. Nucleic Acids Res 32: 1792–1797 13. Kumar et al. Guttikonda SK et al (2010) Whole genome co-expression analysis of soybean cytochrome P450 genes identifies nodulationspecific P450 monooxygenases. FEBS J 278:181 9. Analysis of 727 Cytochrome P450 genes and pseudogenes from a monocot and a dicot. Phytochem Rev 5:193–204 16. Acknowledgments The authors thank to Prof. Earlier studies speculate that non-A-type CYP genes are ancient than A-type families and their organisation requires more time for gene duplication and rearrangement contributing to their slow evolution [17]. Further. CYP82 and CYP76 families and functional analysis of these paralogs might be of great help in understanding the oil and terpenoid metabolism in castor. Peterson D. Peterson N.500) [17] and flax (334/47. Groeneboer S. Reddy TP. Functional characterization of this gene is essential to confirm the same. Nat Biotechnol 28:951–956 7. London. About 83 % of both A-type and non-A-type CYP genes are split genes. while remaining 17 % sans introns indicate that the ancestral CYP gene contains intron(s). type suggesting the rapid expansion of A-type CYP genes. Singh D (1976) Castor . Ming R. presence of orthologs.) and Jatropha curcas L. Nelson DR. T. the remaining CYP genes in Arabidopsis are comparable to the number of CYP genes in castor. Prevalence of single intron containing genes observed in A-type (68 %) over non-A-type (9 %) support the possibility of rapid evolution of A-type genes. etc. CYP87. Nei M. Conclusion Castor genome disclosed 210 putative CYP coding genes grouped into 10 clans consisting of 45 families represented by 77 subfamilies. Nelson DR (2006) Plant cytochrome P450s from moss to poplar. Distinct projection of RcCYPN gene of castor from CYP74 family genes in the phylogenetic tree. Soetaert W (2011) The role of cytochrome P450 monooxygenases in microbial fatty acid metabolism. Saerens K.900) [18]. 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J Ethnopharmacol 5:117–137 6. Plant J 66:194–211 11. 710 and 711 in green algae [15] confirms their ancient nature. a three-fold representation of phase 2 introns in nonA-type (19 %) over A-type (6 %) might contribute to the slower expansion of non-A-type genes. Besides the presence of multiple introns. Extensive amplification of genes was observed in CYP716. CYP87. CYP51. However. After excluding CYP702 and CYP708 genes which are specific to model plant. Van Bogaert IN. grape (315/30. Jones KM.. FEBS J 278:206–221 12. castor CYP716. morphological differences observed between the polymorphic forms were attributed to differences in genes. Scarpa A. Nelson DR.654) [15]. Orvis J. Trop Plant Biol 1:216–235 17. Department of Genetics. Cell Mol Life Sci 58:737–747 8. Anzenbacherova´ E (2001) Cytochrome P450s and metabolism of xenobiotics. CYP82 and CYP76 families recorded extensive amplification of genes plausibly pertaining to oil and terpenoid metabolism. Werck-Reichhart D. About 83 % of CYP genes are split genes indicating their origin from an ancestral CYP gene with intron. About *66 % of the introns of castor CYP genes exhibited zero phase suggesting the plausible evolution of split genes by the inclusion of functional domain coding sequences into single transcription unit.

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