水稻IDD基因家族的全基因组鉴定及综合分析

doi:10.16819/j.1001-7216.2024.240101

摘要/Abstract

摘要：

【目的】明确植物特异转录因子OsIDD基因家族成员特征特性，验证其表达模式，为深入研究水稻IDD基因家族生物学功能奠定基础。【方法】通过Pfam鉴定与明确水稻IDD家族成员，对其核酸与蛋白序列特征、蛋白结构、系统发育与共线性、启动子元件进行综合分析；通过RT-PCR技术验证OsIDD基因在各组织、不同激素处理及不同非生物胁迫下的表达模式。【结果】发现OsIDD家族具有15个成员，为3个亚家族，在染色体上呈不均匀分布，N端均具有核定位信号及高度保守的ID结构域。多重序列比对及motif分析表明其在各个类型中相对保守，与玉米IDD基因家族共线性分析表明两个物种间具有高度同源性。芯片及RT-PCR分析发现OsIDD主要在穗及发育胚乳中表达量较高，其启动子序列均包含激素/胁迫响应顺式元件。【结论】本研究对15个OsIDD成员进行了综合分析，OsIDD结构保守，与ZmIDD共线性强，主要在穗及发育胚乳组织中高表达，可能参与不同激素及多种非生物逆境胁迫响应过程。

关键词: 水稻, IDD基因家族, 综合分析, 表达模式分析

Abstract:

【Objective】To clarify the characteristics and verify the expression patterns of IDD gene family members in rice, which are plant-specific transcription factors, in order to further investigate the biological functions of OsIDD.【Method】The IDD family members in rice were identified using Pfam, followed by a comprehensive analysis of their nucleic acid and protein sequence features, protein structure, phylogeny, collinearity, and promoter elements. RT-PCR was employed to verify the expression patterns of the OsIDD gene in various tissues, under different hormone treatments, and in response to various abiotic stresses.【Result】The OsIDD family consists of 15 members falling into three subfamilies, which are unevenly distributed across the chromosomes. All members possess nuclear localization signals and highly conserved ID structural domains at the N-terminus. Multiple sequence alignment and motif analysis indicate that these sequences are relatively conserved among different types. Collinearity analysis with the ZmIDD gene family reveals a high degree of homology between the two species. Microarray and RT-PCR analyses showed that OsIDD is predominantly expressed in the panicle and developing endosperm, with its promoter sequences containing hormone and stress-responsive cis-elements.【Conclusion】A comprehensive analysis of the 15 OsIDD members demonstrated structural conservation, strong collinearity with ZmIDD, and high expression levels in panicle and developing endosperm. These findings suggest that OsIDD may be involved in the response to different hormones and various abiotic stresses.

Key words: rice, IDD gene family, comprehensive analysis, expression pattern analysis

钟智慧, 秦璐, 黎志力, 杨珍, 贺晓鹏, 蔡怡聪. 水稻IDD基因家族的全基因组鉴定及综合分析[J]. 中国水稻科学, 2024, 38(6): 638-652.

ZHONG Zhihu, QIN Lu, LI Zhili, YANG Zhen, HE Xiaopeng, CAI Yicong. Genome-wide Identification and Comprehensive Analysis of IDD Gene Family in Rice[J]. Chinese Journal OF Rice Science, 2024, 38(6): 638-652.

图/表 14

参考文献 39

[1]	孙燕, 苟德明, 李文鑫. C₂H₂型锌指蛋白研究进展[J]. 生命的化学, 2001 (6): 473-475.
	Sun Y, Gou D M, Li W X. Advances in the study of C₂H₂ type zinc finger proteins[J]. Chemistry of Life, 2001(6): 473-475. (in Chinese with English abstract)
[2]	Macpherson S, Larochelle M, Turcotte B. A fungal family of transcriptional regulators: the zinc cluster proteins[J]. Microbiology and Molecular Biology Reviews, 2006, 70(3): 583-604.
[3]	Lee M S, Gippert G P, Soman K V, Case D A, Wright P E. Three-dimensional solution structure of a single zinc finger DNA-binding domain[J]. Science, 1989, 245(4918): 635-637.
[4]	Párraga G, Horvath S J, Eisen A, Taylor W E, Hood L, Young E T, Klevit R E. Zinc-dependent structure of a single-finger domain of yeast ADR1[J]. Science, 1988, 241(4872): 1489-1492.
[5]	Xu Q K, Yu H P, Xia S S, Cui Y J, Yu X Q, Liu H, Zeng D L, Hu J, Zhang Q, Gao Z Y, Zhang G H, Zhu L, Shen L, Guo L B, Rao Y C, Qian Q, Ren D Y. The C₂H₂ zinc-finger protein LACKING RUDIMENTARY GLUME 1 regulates spikelet development in rice[J]. Science Bulletin, 2020, 65(9): 753-764.
[6]	侯思宇, 孙朝霞, 郭彬, 王玉国, 李贵全, 韩渊怀. 大豆两个C₂H₂型转录因子基因序列特征及表达分析[J]. 植物生理学报, 2014, 50(5): 665-674.
	Hou S Y, Sun Z X, Guo B, Wang Y G, Li G Q, Han Y H. Cloning and expression analysis of two C₂H₂ transcription factors in soybean[J]. Plant Physiology Journal, 2014, 50(5): 665-674. (in Chinese with English abstract)
[7]	孟繁君, 陈明, 徐长营, 朴秀吉, 汪可心, 葛善欣, 金玄吉. 玉米C₂H₂型锌指蛋白基因ZFP225的鉴定、生物信息学分析与克隆[J]. 作物杂志, 2014 (1): 49-53.
	Meng F J, Chen M, Xu C Y, Piao X J, Wang K X, Ge S X, Jin X J. Identification, bioinformatic analysis and cloning of type C₂H₂ zinc finger protein gene ZFP225 in maize[J]. Crops, 2014(1): 49-53. (in Chinese with English abstract)
[8]	Sakamoto H, Araki T, Meshi T, Iwabuchi M. Expression of a subset of the Arabidopsis Cys(2)/His(2)-type zinc-finger protein gene family under water stress[J]. Gene, 2000, 248(1-2): 23-32.
[9]	Park S J, Kim S L, Lee S, Je B I, Piao H L, Park S H, Kim C M, Ryu C H, Park S H, Xuan Y H, Colasanti J, An G, Han C D. Rice Indeterminate 1 (OsId1) is necessary for the expression of Ehd1 (Early heading date 1) regardless of photoperiod[J]. The Plant Journal, 2008, 56(6): 1018-1029.
[10]	Matsubara K, Yamanouchi U, Wang Z X, Minobe Y, Izawa T, Yano M. Ehd2, a rice ortholog of the maize INDETERMINATE1 gene, promotes flowering by up-regulating Ehd1[J]. Plant Physiology, 2008, 148(3): 1425-1435.
[11]	Colasanti J, Tremblay R, Wong A Y, Coneva V, Kozaki A, Mable B K. The maize INDETERMINATE1 flowering time regulator defines a highly conserved zinc finger protein family in higher plants[J]. BMC Genomics, 2006, 7: 158.
[12]	Colasanti J, Yuan Z, Sundaresan V. The indeterminate gene encodes a zinc finger protein and regulates a leaf-generated signal required for the transition to flowering in maize[J]. Cell, 1998, 93(4): 593-603.
[13]	Wu C Y, You C J, Li C S, Long T, Chen G X, Byrne M E, Zhang Q F. RID1, encoding a Cys2/His2-type zinc finger transcription factor, acts as a master switch from vegetative to floral development in rice[J]. Proceedings of the National Academy of Sciences of the United States of America, 2008, 105(35): 12915-12920.
[14]	Coneva V, Guevara D, Rothstein S J, Colasanti J. Transcript and metabolite signature of maize source leaves suggests a link between transitory starch to sucrose balance and the autonomous floral transition[J]. Journal of Experimental Botany, 2012, 63(2): 5079-5092.
[15]	Sakamoto H, Maruyama K, Sakuma Y, Meshi T, Iwabuchi M, Shinozaki K, Yamaguchi-Shinozaki K. Arabidopsis Cys2/His2-type zinc-finger proteins function as transcription repressors under drought, cold, and high-salinity stress conditions[J]. Plant Physiology, 2004, 136(1): 2734-2746.
[16]	Gontarek B C, Neelakandan A K, Wu H, Becraft P W. NKD transcription factors are central regulators of maize endosperm development[J]. The Plant Cell, 2016, 28(12): 2916-2936.
[17]	Coelho C P, Huang P, Lee D Y, Brutnell T P. Making roots, shoots, and seeds: IDD gene family diversification in plants[J]. Trends in Plant Science, 2018, 23(1): 66-78.
[18]	Morita M T, Sakaguchi K, Kiyose S I, Taira K, Kato T, Nakamura M, Tasaka M. A C₂H₂‐type zinc finger protein, SGR5, is involved in early events of gravitropism in Arabidopsis inflorescence stems[J]. The Plant Journal, 2006, 47(4): 619-628.
[19]	Cui D Y, Zhao J B, Jing Y J, Fan M Z, Liu J, Wang Z C, Xin W, Hu Y X. The Arabidopsis IDD14, IDD15, and IDD16 cooperatively regulate lateral organ morphogenesis and gravitropism by promoting auxin biosynthesis and transport[J]. PLoS Genetics, 2013, 9(9): e1003759.
[20]	Feurtado J A, Huang D, Wicki-Stordeur L, Hemstock L E, Potentier M S, Tsang E W, Cutler A J. The Arabidopsis C₂H₂ zinc finger INDETERMINATE DOMAIN1/ ENHYDROUS promotes the transition to germination by regulating light and hormonal signaling during seed maturation[J]. The Plant Cell, 2011, 23(5): 1772-1794.
[21]	Patterson K, Cakmak T, Cooper A, Lager I, Rasmusson A G, Escobar M A. Distinct signalling pathways and transcriptome response signatures differentiate ammonium- and nitrate-supplied plants[J]. Plant, Cell & Environment, 2010, 33(9): 1486-1501.
[22]	Seo P J, Ryu J, Kang S K, Park C M. Modulation of sugar metabolism by an INDETERMINATE DOMAIN transcription factor contributes to photoperiodic flowering in Arabidopsis[J]. The Plant Journal, 2011, 65(3): 418-429.
[23]	Matsubara K, Yamanouchi U, Wang Z X, Minobe Y, Izawa T, Yano M. Ehd2, a rice ortholog of the maize INDETERMINATE1 gene, promotes flowering by up-regulating Ehd1[J]. Plant Physiology, 2008, 148(3): 1425-1435.
[24]	Deng L, Li L M, Zhang S, Shen J Q, Li S B, Hu S F, Peng Q, Xiao J H, Wu C Y. Suppressor of rid1 (SID1) shares common targets with RID1 on florigen genes to initiate floral transition in rice[J]. PLOS Genetics, 2017, 13(2): e1006642.
[25]	Dou M Z, Cheng S, Zhao B T, Xuan Y H, Shao M L. The Indeterminate Domain Protein ROC1 regulates chilling tolerance via activation of DREB1B/CBF1 in rice[J]. International Journal of Molecular Sciences, 2016, 17(3): 233.
[26]	Liu J M, Park S J, Huang J, Lee E J, Xuan Y H, Je B I, Kumar V, Priatama R A, Raj K V, Kim S H, Min M K, Cho J H, Kim T H, Chandran A K, Jung K H, Takatsuto S, Fujioka S, Han C D. Loose Plant Architecture1 (LPA1) determines lamina joint bending by suppressing auxin signalling that interacts with C-22-hydroxylated and 6-deoxo brassinosteroids in rice[J]. Journal of Experimental Botany, 2016, 67(6): 1883-1895.
[27]	Wu X R, Tang D, Li M, Wang K J, Cheng Z K. Loose Plant Architecture1, an INDETERMINATE DOMAIN Protein involved in shoot gravitropism, regulates plant architecture in rice[J]. Plant Physiology, 2013, 161(1): 317-329.
[28]	Cui Z B, Xue C Y, Mei Q, Xuan Y H. Malectin Domain Protein Kinase (MDPK) promotes rice resistance to sheath blight via IDD12, IDD13, and IDD14[J]. International Journal of Molecular Sciences, 2022, 23(15): 8214.
[29]	Livak K J, Schmittgen T D. Analysis of relative gene expression data using realtime quantitative pcr and the 2^-△△CT method[J]. Methods, 2001, 25(4): 402-408.
[30]	Malheiros R S P, Costa L C, Ávila R T, Pimenta T M, Teixeira L S, Brito F A L, Zsögön A, Araújo W L, Ribeiro D M. Selenium downregulates auxin and ethylene biosynthesis in rice seedlings to modify primary metabolism and root architecture[J]. Planta, 2019, 250(1): 333-345.
[31]	Fu Z Z, Yu J, Cheng X W, Zong X, Xu J, Chen M J, Li Z Y, Zhang D B, Liang W Q. The rice Basic Helix-Loop- Helix Transcription Factor TDR INTERACTING PROTEIN2 is a central switch in early anther development[J]. The Plant Cell, 2014, 26(4): 1512-1524.
[32]	Ko S S, Li M J, Sun-Ben Ku M, Ho Y C, Lin Y J, Chuang M H, Hsing H X, Lien Y C, Yang H T, Chang H C, Chan M T. The bHLH142 transcription factor coordinates with TDR1 to modulate the expression of EAT1 and regulate pollen development in rice[J]. The Plant Cell, 2014, 26(6): 2486-2504.
[33]	杜逸凡, 方希林, 曾红丽, 于玉凤, 张露倩, 王悦. 外源多效唑, ABA, 2, 4-D和NAA对水稻秧苗生长特性的影响[J]. 分子植物育种, 2020, 18(8): 2687-2694.
	Du Y F, Fang X L, Zeng H L, Yu Y F, Zhang L Q, Wang Y. Effects of exogenous Paclobutrazol, ABA, 2, 4-D, and NAA on growth characteristics of rice seedlings[J]. Molecular Plant Breeding, 18(8): 2687-2694. (in Chinese with English abstract)
[34]	Huang S Y, Liu M M, Chen G L, Si F F, Fan F F, Guo Y, Yuan L, Yang F, Li S Q. Favorable QTLs from Oryza longistaminata improve rice drought resistance[J]. BMC Plant Biology, 2022, 22(1): 136.
[35]	Song Y, Zhang C J, Ge W N, Zhang Y F, Burlingame A L, Guo Y. Identification of NaCl stress-responsive apoplastic proteins in rice shoot stems by 2D-DIGE[J]. Journal of Proteomics, 2011, 74(7): 1045-1067.
[36]	Kozaki A, Hake S, Colasanti J. The maize ID1 flowering time regulator is a zinc finger protein with novel DNA binding properties[J]. Nucleic Acids Research, 2004, 32(5): 1710-1720.
[37]	Seo P J, Kim M J, Ryu J Y, Jeong E Y, Park C M. Two splice variants of the IDD14 transcription factor competitively form nonfunctional heterodimers which may regulate starch metabolism[J]. Nature Communications, 2011, 2: 303.
[38]	Sun Q, Li T Y, Li D D, Wang Z Y, Li S, Li D P, Han X, Liu J M, Xuan Y H. Overexpression of Loose Plant Architecture 1 increases planting density and resistance to sheath blight disease via activation of PIN‐FORMED 1a in rice[J]. Plant Biotechnology Journal, 2019, 17(5): 855-857.
[39]	Lu Y Z, Feng Z, Meng Y L, Bian L Y, Xie H, Mysore K S, Liang J S. SLENDER RICE1 and Oryza sativa INDETERMINATE DOMAIN 2 regulating OsmiR396 are involved in stem elongation[J]. Plant Physiology, 2020, 182(4): 2213-2227.

引物名称 Primer name	正向引物序列 Forward sequence (5’-3’)	反向引物序列 Revers sequence(5’-3’)
Actin-RT	CGGGAAATTGTGAGGGACAT	AGGAAGGCTGGAAGAGGACC
OsID1-RT	CTCTTCTCCAGGAAGGACAGCC	GTAGTAGTGATGCTGCTGCTGTTG
OsIDD1-RT	GTTCTGGTCGCGCTGGAAC	GACACAATCATTAGGAGCGGCA
OsIDD2-RT	CGGCAACCCAGATCCTGAG	CTGGTCTCGCTGGAACCC
OsIDD3-RT	GGAAACCCAAATCCAGATGCG	CCGCTGGAACCCCTTGTTG
OsIDD4-RT	CCTTACCAGACCCGGACGC	GTTCTGCTCCCGCTGGAAC
OsIDD5-RT	GGATGAGCAAGAGGTGGTTG	ATCCAGCAGGTTTGAGTCCT
OsIDD6-RT	CCCGGGAATCCAAATCCTGATGC	CTGCTCCCTCTGGAACCCC
OsIDD7-RT	GGAAACCCAGATCCAGATGTTGAG	GGAGGTTCTGGTCTCTCTGGAAG
OsIDD8-RT	CGGCACACCAGATCCGG	CAGGTTCTGGTCTCGCTGG
OsIDD9-RT	GTCTTCTCCAGGCGCGAC	CAGAGCGGAGGTGACCG
OsIDD10-RT	CAACCAACCCAAACCCGGAC	GGTTCTGCTCCCGCTGG
OsIDD11-RT	GCAACCCAGACCCGGAAG	CTGCAGGTTCTGGTCCCTC
OsIDD12-RT	CAGATCACCTGCTACAGCTG	TGACGACGGACGAGACGTA
OsIDD13-RT	GCACGCCAGACCCGGAC	CTGCAGGTTCTGGTCTCGC
OsIDD14-RT	GCGTCTTCTCCCGAGTGG	GGATGACCGGCAGCGTC

引物名称 Primer name	正向引物序列 Forward sequence (5’-3’)	反向引物序列 Revers sequence(5’-3’)
Actin-RT	CGGGAAATTGTGAGGGACAT	AGGAAGGCTGGAAGAGGACC
OsID1-RT	CTCTTCTCCAGGAAGGACAGCC	GTAGTAGTGATGCTGCTGCTGTTG
OsIDD1-RT	GTTCTGGTCGCGCTGGAAC	GACACAATCATTAGGAGCGGCA
OsIDD2-RT	CGGCAACCCAGATCCTGAG	CTGGTCTCGCTGGAACCC
OsIDD3-RT	GGAAACCCAAATCCAGATGCG	CCGCTGGAACCCCTTGTTG
OsIDD4-RT	CCTTACCAGACCCGGACGC	GTTCTGCTCCCGCTGGAAC
OsIDD5-RT	GGATGAGCAAGAGGTGGTTG	ATCCAGCAGGTTTGAGTCCT
OsIDD6-RT	CCCGGGAATCCAAATCCTGATGC	CTGCTCCCTCTGGAACCCC
OsIDD7-RT	GGAAACCCAGATCCAGATGTTGAG	GGAGGTTCTGGTCTCTCTGGAAG
OsIDD8-RT	CGGCACACCAGATCCGG	CAGGTTCTGGTCTCGCTGG
OsIDD9-RT	GTCTTCTCCAGGCGCGAC	CAGAGCGGAGGTGACCG
OsIDD10-RT	CAACCAACCCAAACCCGGAC	GGTTCTGCTCCCGCTGG
OsIDD11-RT	GCAACCCAGACCCGGAAG	CTGCAGGTTCTGGTCCCTC
OsIDD12-RT	CAGATCACCTGCTACAGCTG	TGACGACGGACGAGACGTA
OsIDD13-RT	GCACGCCAGACCCGGAC	CTGCAGGTTCTGGTCTCGC
OsIDD14-RT	GCGTCTTCTCCCGAGTGG	GGATGACCGGCAGCGTC

名称 Name	染色体 Chr.	基因长度 Gene length	氨基酸数目 No. of amino acids	分子量 Molecular weight	理论等电点 Theoretical pI	蛋白质不稳定指数 Instability index	脂溶指数 Aliphatic index	亲水性平均系数 Grand average of hydropathicity	亚细胞定位预测 Predicted subcellular localization
OsID1	10	1230	409	44267.44	8.04870033	51.19	51.19	−0.467	细胞核Nucleus
OsIDD1	3	1659	552	57540.01	8.55090046	75.58	75.58	−0.486	细胞核Nucleus
OsIDD2	1	1464	487	51815.90	8.60980034	48.96	48.96	−0.527	细胞核Nucleus
OsIDD3	9	1608	535	54893.05	9.41469955	46.65	46.65	−0.460	细胞核Nucleus
OsIDD4	2	1848	615	63597.71	8.63129997	46.29	46.29	−0.497	细胞核Nucleus
OsIDD5	7	1902	633	67636.09	6.01849985	55.08	55.08	−0.567	细胞核Nucleus
OsIDD6	8	1659	552	57262.74	9.38479996	49.81	49.81	−0.500	细胞核Nucleus
OsIDD7	2	1479	492	54491.89	7.31449986	59.23	59.23	−0.697	细胞核Nucleus
OsIDD8	1	1488	495	51394.20	8.89400005	49.83	49.83	−0.293	细胞核Nucleus
OsIDD9	1	1431	476	50107.14	7.95529985	52.62	52.62	−0.265	细胞核Nucleus
OsIDD10	4	1842	613	63814.37	9.93620014	52.40	52.4	−0.476	细胞核Nucleus
OsIDD11	1	1614	537	57410.90	7.97130013	62.67	62.67	−0.703	细胞核Nucleus
OsIDD12	8	1602	533	56426.28	9.51509953	62.47	62.47	−0.467	细胞核Nucleus
OsIDD13	9	1515	504	53574.19	8.50090027	62.67	62.67	−0.459	细胞核Nucleus
OsIDD14	3	1317	438	45959.66	10.12440010	70.46	70.46	−0.509	细胞核Nucleus

名称 Name	染色体 Chr.	基因长度 Gene length	氨基酸数目 No. of amino acids	分子量 Molecular weight	理论等电点 Theoretical pI	蛋白质不稳定指数 Instability index	脂溶指数 Aliphatic index	亲水性平均系数 Grand average of hydropathicity	亚细胞定位预测 Predicted subcellular localization
OsID1	10	1230	409	44267.44	8.04870033	51.19	51.19	−0.467	细胞核Nucleus
OsIDD1	3	1659	552	57540.01	8.55090046	75.58	75.58	−0.486	细胞核Nucleus
OsIDD2	1	1464	487	51815.90	8.60980034	48.96	48.96	−0.527	细胞核Nucleus
OsIDD3	9	1608	535	54893.05	9.41469955	46.65	46.65	−0.460	细胞核Nucleus
OsIDD4	2	1848	615	63597.71	8.63129997	46.29	46.29	−0.497	细胞核Nucleus
OsIDD5	7	1902	633	67636.09	6.01849985	55.08	55.08	−0.567	细胞核Nucleus
OsIDD6	8	1659	552	57262.74	9.38479996	49.81	49.81	−0.500	细胞核Nucleus
OsIDD7	2	1479	492	54491.89	7.31449986	59.23	59.23	−0.697	细胞核Nucleus
OsIDD8	1	1488	495	51394.20	8.89400005	49.83	49.83	−0.293	细胞核Nucleus
OsIDD9	1	1431	476	50107.14	7.95529985	52.62	52.62	−0.265	细胞核Nucleus
OsIDD10	4	1842	613	63814.37	9.93620014	52.40	52.4	−0.476	细胞核Nucleus
OsIDD11	1	1614	537	57410.90	7.97130013	62.67	62.67	−0.703	细胞核Nucleus
OsIDD12	8	1602	533	56426.28	9.51509953	62.47	62.47	−0.467	细胞核Nucleus
OsIDD13	9	1515	504	53574.19	8.50090027	62.67	62.67	−0.459	细胞核Nucleus
OsIDD14	3	1317	438	45959.66	10.12440010	70.46	70.46	−0.509	细胞核Nucleus

基因名称 Gene name	α-螺旋 α-helix	延伸链 Extended strand	β-转角 β-turn	无规则卷曲 Random coil
OsID1	92 (22.49%)	89 (21.76%)	33 (8.07%)	195 (47.68%)
OsIDD1	78 (14.13%)	66 (11.96%)	31 (5.62%)	377 (68.30%)
OsIDD2	149 (30.60%)	42 (8.62%)	32 (6.57%)	264 (54.21%)
OsIDD3	97 (18.13%)	71 (13.27%)	30 (5.61%)	337 (62.99%)
OsIDD4	182 (29.59%)	77 (12.52%)	48 (7.80%)	308 (50.08%)
OsIDD5	132 (20.85%)	97 (15.32%)	50 (7.90%)	354 (55.92%)
OsIDD6	150 (27.17%)	87 (15.76%)	48 (8.70%)	267 (48.37%)
OsIDD7	99 (20.12%)	48 (9.76%)	16 (3.25%)	329 (66.87%)
OsIDD8	147 (29.70%)	64 (12.93%)	42 (8.48%)	242 (48.89%)
OsIDD9	112 (23.53%)	68 (14.29%)	45 (9.45%)	251 (52.73%)
OsIDD10	172 (28.06%)	82 (13.38%)	37 (6.04%)	322 (52.53%)
OsIDD11	195 (36.31%)	58 (10.80%)	46 (8.57%)	238 (44.32%)
OsIDD12	200 (37.52%)	69 (12.95%)	33 (6.19%)	231 (43.34%)
OsIDD13	225 (44.64%)	40 (7.94%)	13 (2.58%)	226 (44.84%)
OsIDD14	190 (43.38%)	41 (9.36%)	35 (7.99%)	172 (39.27%)