甜瓜SLAF图谱构建及果实相关性状QTL分析

doi:10.3864/j.issn.0578-1752.2021.19.014

摘要/Abstract

摘要：

【目的】通过对甜瓜果实相关性状进行QTL定位及候选基因分析,为甜瓜品质育种及基因挖掘与功能验证提供理论基础。【方法】利用薄皮甜瓜1244为母本、厚皮甜瓜M5为父本配置杂交组合,结合SFLA测序技术开发分子标签,构建高密度遗传图谱,以F_2:3表型数据为基础,采用Mapqtl进行QTL定位分析。【结果】获得了380 446 Mreads（83.12 Gb）数据,测序平均Q30为93.59%,平均GC含量为36.87%;开发了112 844个SLAF标签,3 274 879个SNP;构建了12个连锁群,共计10 596个上图标记,总图距为1 383.88 cM,标记间平均距离为0.13 cM,上图标记完整度平均为99.92%。将控制甜瓜果面沟性状基因（fst）定位在第11条染色体末端Marker 1993423（62.18）—Marker 1998820（63.05）,覆盖基因组0.72 Mb,包含33个候选基因;控制甜瓜果皮花纹基因（fpp）定位在第2条染色体Marker 459584（90.91）—Marker 459446（90.91）,覆盖基因组0.08 Mb,包含5个候选基因,其中MELO3C026292（1-脱氧-D-木酮糖-5-磷酸还原异构酶）可能为控制果皮花纹的候选基因;同时检测到甜瓜果皮底色1个QTL位点fpc,位于第7条染色体Marker 1229174（7.14）—Marker 1229973（7.14）,贡献率为9.9%;检测到果型指数1个QTL位点fs9.1,位于第9条染色体Marker 1705671（76.19）—Marker 1705915（79.23）,贡献率为7.6%;在第1、2、6、7、10染色体检测到单果重相关6个QTL位点（sfw1.1、sfw2.1、sfw2.2、sfw6.1、sfw7.1、sfw10.1）,贡献率在3.1%—17.0%,LOD值介于3.0—5.6。【结论】将甜瓜果面沟、果皮花纹基因分别定位在第11和第2条染色体,分别获得33个和5个候选基因;检测到1个果皮底色QTL位点、1个果型指数QTL位点和6个单果重QTL位点。

关键词: 甜瓜, SLAF测序, 遗传图谱构建, 果实性状, QTL分析

Abstract:

【Objective】 Through the QTL mapping and candidate gene analysis of muskmelon (Cucumis melo L.) fruit-related traits, so as to provide a theoretical basis for muskmelon quality breeding, gene mapping and functional verification. 【Method】 The F₁ plant was made cross by 1244 as female parent with thin-skinned melon and M5 with thick-skinned melon as male parent, combined with SFLA sequencing technology to develop molecular tags and construct high-density genetic maps. Based on F_2:3 phenotype data, Mapqtl analysis method was used to detect QTL location. 【Result】 Totally 380 446 Mreads (83.12 Gb) data were obtained, the average Q30 and GC content was 93.59% and 36.87%, respectively; totally 112 844 SLAF tags and 3 274 879 SNPs were obtained; a genetic map contained 12 linkage groups with a total of 10 596 markers were constructed. The genetic map spanned 1 383.88 cM with the average distance was 0.13 cM between markers, and totally 99.92% developed markers were mapped. The gene controling fruit sutures (fst) of melon was located at the end of chromosome 11, between Marker 1993423 (62.18) and Marker 1998820 (63.05), covering 0.72 Mb of the genome, and containing 33 candidate genes. The gene for pericarp pattern (fpp) was located at chromosome 2 between Marker 459584 (90.91) and Marker 459446 (90.91) which covered the genome of 0.08 Mb and contained 5 candidate genes, among them MELO3C026292 (1-deoxy-D-xylulose-5-phosphate reductoisomerase) might be the candidate gene for pericarp pattern. At the same time, QTL locus of pericarp color (fpc) was detected which located between Marker 1229174 (7.14) and Marker 1229973 (7.14) on the chromosome 7, with the contribution rate was 9.9%. One QTL locus of fruit shape (fs) fs9.1 was detected between Marker 1705671 (76.19) and Marker 1705915 (79.23) on chromosome 9 with a contribution rate of 7.6%. Six QTL loci related to single fruit weight (sfw1.1, sfw2.1, sfw2.2, sfw6.1, sfw7.1, sfw10.1) were detected on chromosomes 1, 2, 6, 7, and 10 with the contribution rate were between 3.1% to 17%, and the LOD value were between 3.0 to 5.6. 【Conclusion】The fruit sutures and pericarp pattern genes were located on the chromosomes11 and 2, and then 33 and 5 candidate genes were obtained, respectively; One QTL for pericarp color, one QTL for fruit shape and 6 QTLs for single fruit weight were also detected.

Key words: melon, SLAF sequencing, genetic map construction, fruit related traits, QTL analysis

王岭,才羿,王桂超,王迪,盛云燕. 甜瓜SLAF图谱构建及果实相关性状QTL分析[J]. 中国农业科学, 2021, 54(19): 4196-4206.

WANG Ling,CAI Yi,WANG GuiChao,WANG Di,SHENG YunYan. Specific Length Amplified Fragment (SFLA) Sequencing Mapping Construction and QTL Analysis of Fruit Related Traits in Muskmelon[J]. Scientia Agricultura Sinica, 2021, 54(19): 4196-4206.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.chinaagrisci.com/CN/10.3864/j.issn.0578-1752.2021.19.014

https://www.chinaagrisci.com/CN/Y2021/V54/I19/4196

图/表 9

图1

表1

图2

表2

表3

表4

图3

图4

表5

参考文献 32

[1]	ZHAO G W, LIAN Q, ZHANG Z H, FU Q S, HE Y H, MA S W, RUGGIERI V, MONFORTE A J, WANG P Y, JULCA I, WANG H S, LIU J P, XU Y, WANG R Z, JI J B, XU Z H, KONG W H, ZHONG Y, SHANG J L, PEREIRA L, et al. A comprehensive genome variation map of melon identifies multiple domestication events and loci influencing agronomic traits. Nature Genetics, 2019, 51:1607-1615. doi: 10.1038/s41588-019-0522-8
[2]	高鹏, 刘识, 崔浩楠, 张泰峰, 王学征, 刘宏宇, 朱子成, 栾非时. 甜瓜基因组学、功能基因定位及基因工程育种研究进展. 园艺学报, 2020, 47(9):1827-1844.
	GAO P, LIU S, CUI H N, ZHANG T F, WANG X Z, LIU H Y, ZHU Z C, LUAN F S. Research progress of melon genomics, functional gene mapping and genetic engineering. Acta Horticulturae Sinica, 2020, 47(9):1827-1844. (in Chinese)
[3]	刘柳. 甜瓜果皮条纹的遗传分析及其决定基因的精细定位[D]. 哈尔滨: 东北农业大学, 2019.
	LIU L. Genetic analysis of rind stripe and fine mapping of the determinate gene in melon (Cucumis melo L.)[D]. Harbin: Northeast Agricultural University, 2019. (in Chinese)
[4]	吕建春. 薄皮甜瓜(Cucumis melo L. var. chinensis Pangalo)果皮花斑的遗传分析及基因定位[D]. 北京: 中国农业科学院, 2018.
	LÜ J C. Inheritance and gene mapping of spotted trait in melon (Cucumis melo L. var. chinensis Pangalo)[D]. Beijing: Chinese Academy of Agricultural Sciences, 2018. (in Chinese)
[5]	欧点点. 甜瓜果皮颜色遗传分析及绿色果皮基因定位[D]. 北京:中国农业科学院, 2019.
	OU D D. Genetic Analysis and Gene Mapping for Green Peel Color in Melon (Cucumis melo L.)[D]. Beijing: Chinese Academy of Agricultural Sciences, 2019. (in Chinese)
[6]	GALPAZ N, GONDA I, SHEM-TOV D, BARAD O, TZURI G, LEV S, FEI Z J, XU Y M, MAO L Y, JIAO C, HAREL-BEJA R, DORON-FAIGENBOIM A, TZFADIA O, MEIR E B A, SA'AR U, FAIT A, HALPERIN E, KENIGSWALD M, FALLIK E, KOL N L G, et al. Deciphering genetic factors that determine melon fruit-quality traits using RNA seq-based high-resolution QTL and eQTL mapping. Journal of Molecular Cell Biology, 2018, 94(1):169-191.
[7]	杨光华, 范荣, 杨小锋, 侯军亮, 袁士臣, 曹明, 王学林, 李劲松. 甜瓜果实颜色3个质量性状基因的定位. 园艺学报, 2014, 41(5):898-906.
	YANG G H, FAN R, YANG X F, HOU J L, YUAN S C, CAO M, WANG X L, LI J S. Construction of a highly dense genetic map using SNP and mapping of three qualitative traits in Cucumis melo. Acta Horticulturae Sinica, 2014, 41(5):898-906. (in Chinese)
[8]	GUR A, TZURI G, MEIR A, SA'AR U, PORTNOY V, KATZIR N, SCHAFFER A A, LI L, BURGER J, TADMOR Y. Genome-wide linkage-disequilibrium mapping to the candidate gene level in melon (Cucumis melo). Scientific Reports, 2017, 7:9770. doi: 10.1038/s41598-017-09987-4
[9]	FEDER A, BURGER J, GAO S, LEWINSOHN E, KATZIR N, SCHAFFER A A, MEIR A, DAVIDOVICH-RIKANATI R, PORTNOY V, GAL-ON A, FEI Z J, KASHI Y, TADMOR Y. Focus on metabolism: A kelch domain-containing F-Box coding gene negatively regulates flavonoid accumulation in muskmelon. Plant Physiology, 2015, 169(3):1714-1726.
[10]	邱果. 甜瓜果面沟相关基因定位与相关性分析[D]. 哈尔滨: 东北农业大学, 2018.
	QIU G. Location of surface groove related gene and correlation analysis in melon[D]. Harbin: Northeast Agricultural University, 2018. (in Chinese)
[11]	CHANG C W, WANG Y H, TUNG C W. Genome-wide single nucleotide polymorphism discovery and the construction of a high- density genetic map for melon (Cucumis melo L.) using genotyping- by-sequencing. Frontiers in Plant Science, 2017, 8:125.
[12]	WU S, ZHANG B Y, KEYHANINEJAD N, RODRIGUEZ G R, KIM H J, CHAKRABARTI M, ILLA-BERENGUER E, TAITANO N K, GONZALO M J, DIAZ A, PAN Y P, LEISNERC P, HALTERMAN D, BUELL C R, WENG Y Q, JANSKY S H, VAN ECK H, WILLEMSEN J, MONFORTE A J, MEULIA T, VAN DER KNAAP E. A common genetic mechanism underlies morphological diversity in fruits and other plant organs. Nature Communications, 2018, 9:4734. doi: 10.1038/s41467-018-07216-8
[13]	王晨晖, 栾非时, 高鹏, 刘识, 解志强. 菜瓜×马泡瓜遗传连锁图谱构建及果实相关性状QTL分析. 园艺学报, 2019, 46(12):2347-2358.
	WANG C H, LUAN F S, GAO P, LIU S, XIE Z Q. Construction of melon genetic linkage map and QTL analysis of fruit related traits in snake melon × wild type melon genetic background. Acta Horticulturae Sinica, 2019, 46(12):2347-2358. (in Chinese)
[14]	胡志程, 周梦迪, 吕建春, 付秋实, 王怀松. 甜瓜遗传图谱与基因定位研究进展. 分子植物育种, 2020, 18(7):2290-2295.
	HU Z C, ZHOU M D, LÜ J C, FU Q S, WANG H S. The research progress of genetic maps and genes mapping on melon. Molecular Plant Breeding, 2020, 18(7):2290-2295. (in Chinese)
[15]	GARCIA-MAS J, BENJAK A, SANSEVERINO W, BOURGEOIS M, MIR G, GONZÁLEZ V M, HÉNAFF E, CÂMARA F, COZZUTO L, LOWY E, ALIOTO T, CAPELLA-GUTIÉRREZ S, BLANCA J, CAÑIZARES J, ZIARSOLO P, GONZALEZ-IBEAS D, RODRÍGUEZ-MORENO L, DROEGE M, DU L, ALVAREZ-TEJADO M, et al. The genome of melon (Cucumis melo L.). Proceedings of the National Academy of Sciences of the United States of America, 2012, 109(29):11872-11877.
[16]	栾非时, 矫士琦, 盛云燕, 朱子成. 甜瓜果实相关性状QTL分析. 东北农业大学学报, 2017, 48(3):1-9.
	LUAN F S, JIAO S Q, SHENG Y Y, ZHU Z C. Mapping of QTL for fruit traits in melon. Journal of Northeast Agricultural University, 2017, 48(3):1-9. (in Chinese)
[17]	张学军, 杨文莉, 张永兵, 张健, 郭丽霞, 杨永, 李寐华, 伊鸿平. 采用GBS-seq技术构建甜瓜高密度遗传图谱. 新疆农业科学, 2019(10):1828-1838.
	ZHANG X J, YANG W L, ZHANG Y B, ZHANG J, GUO L X, YANG Y, LI M H, YI H P. Construction of high density genetic map and QTL mapping for downy mildew traits in Cucumis melo ssp. melo. Xinjiang Agricultural Sciences, 2019(10):1828-1838. (in Chinese)
[18]	刘相玉, 张裕舒, 刘柳, 刘识, 高鹏, 王迪, 王学征. 基于CAPS标记的甜瓜单果重相关性状QTL分析. 中国农业科学, 2019, 52(9):1601-1613.
	LIU X Y, ZHANG Y S, LIU L, LIU S, GAO P, WANG D, WANG X Z. The QTL analysis of single fruit weight associated traits in melon based on CAPS markers. Scientia Agricultura Sinica, 2019, 52(9):1601-1613. (in Chinese)
[19]	张肖静, 张凯歌, 朱华玉, 张宇, 胡倩梅, 程思源, 张敏娟, 胡建斌, 杨路明. 甜瓜侧枝相关性状的QTL定位. 园艺学报, 2019, 46(3):519-528.
	ZHANG X J, ZHANG K G, ZHU H Y, ZHANG Y, HU Q M, CHENG S Y, ZHANG M J, HU J B, YANG L M. QTL mapping of lateral branch associated traits in Cucumis melo. Acta Horticulturae Sinica, 2019, 46(3):519-528. (in Chinese)
[20]	TZURI G, ZHOU X J, CHAYUT N, YUAN H, PORTNOY V, MEIR A, SA'AR U, BAUMKOLER F, MAZOUREK M, LEWINSOHN E, FEI Z J, SCHAFFER A A, LI L, BURGER J, KATZIR N, TADMOR Y. A ‘golden’ SNP in CmOr governs the fruit flesh color of melon (Cucumis melo). The Plant Journal, 2015, 82(2):267-279. doi: 10.1111/tpj.2015.82.issue-2
[21]	ZHANG H, LI X M, YU H Y, ZHANG Y B, LI M H, WANG H J, WANG D M, WANG H S, FU Q S, LIU M, JI C M, MA L M, TANG J, LI S, MIAO J S, ZHENG H K, YI H P. A high-quality melon genome assembly provides insights into genetic basis of fruit trait improvement. IScience, 2019, 22:16-27. doi: 10.1016/j.isci.2019.10.049
[22]	DIAZ A, FERGANY M, FORMISANO G, ZIARSOLO P, BLANCA J, FEI Z J, STAUB J E, ZALAPA J E, CUEVAS H E, DACE G, OLIVER M, BOISSOT N, DOGIMONT C, PITRAT M, HOFSTEDE R, VAN KOERT P, HAREL-BEJA R, TZURI G, PORTNOY V, COHEN S, SCHAFFER A, KATZIR N, XU Y, ZHANG H Y, FUKINO N, MATSUMOTO S, GARCIA-MAS J, MONFORTE A J. A consensus linkage map for molecular markers and quantitative trait loci associated with economically important traits in melon (Cucumis melo L.). BMC Plant Biology, 2011, 11(1):111. doi: 10.1186/1471-2229-11-111
[23]	DÍAZ A, ZAROURI B, FERGANY M, EDUARDO I, ALVAREZ J M, PICÓ B, MONFORTE A J. Mapping and introgression of QTL involved in fruit shape transgressive segregation into ‘piel de sapo’ melon (Cucumis melo L.). PLoS ONE, 2014, 9(8):e104188. doi: 10.1371/journal.pone.0104188
[24]	PERPIÑÁ G, ESTERAS C, GIBON Y, MONFORTE A J, PICÓ B. A new genomic library of melon introgression lines in a cantaloupe genetic background for dissecting desirable agronomical traits. BMC Plant Biology, 2016, 16(1):154. doi: 10.1186/s12870-016-0842-0
[25]	RAMAMURTHY R K, WATERS B M. Identification of fruit quality and morphology QTLs in melon (Cucumis melo L.) using a population derived from flexuosus and cantalupensis botanical groups. Euphytica, 2015, 204:163-177. doi: 10.1007/s10681-015-1361-z
[26]	王贤磊, 高兴旺, 李冠, 王惠林, 耿守东, 康锋, 聂祥祥. 甜瓜遗传图谱的构建及果实与种子QTL分析. 遗传, 2011, 33(12):1398-1408.
	WANG X L, GAO X W, LI G, WANG H L, GENG S D, KANG F, NIE X X. Construction of a melon genetic map with fruit and seed QTLs. Hereditas, 2011, 33(12):1398-1408. (in Chinese)
[27]	张宁, 张显, 张勇, 马建祥, 杨小振, 王永琦. 甜瓜果实糖含量相关性状QTL分析. 西北植物学报, 2015, 35(2):252-257.
	ZHANG N, ZHANG X A, ZHANG Y, MA J X, YANG X Z, WANG Y Q. QTL analysis of fruit sugar content correlated traits in melon. Acta Botanica Boreali-Occidentalia Sinica, 2015, 35(2):252-257. (in Chinese)
[28]	张雪娇, 高鹏, 栾非时. 甜瓜果实相关性状QTL分析. 中国蔬菜, 2013(18):35-41.
	ZHANG X J, GAO P, LUAN F S. QTL analysis of fruits related traits in melon(Cucumis melo L.). China Vegetables, 2013(18):35-41. (in Chinese)
[29]	周鹏, 李钱峰, 熊敏, 范晓磊, 赵冬生, 张昌泉, 刘巧泉. DELLA蛋白介导的激素互作调控植物生长发育研究进展. 植物生理学报, 2020, 56(4):661-671.
	ZHOU P, LI Q F, XIONG M, FAN X L, ZHAO D S, ZHANG C Q, LIU Q Q. Advances in DELLA protein-mediated phytohormonal crosstalk in regulation of plant growth and development. Plant Physiology Communications, 2020, 56(4):661-671. (in Chinese)
[30]	PAN Y P, WANG Y H, MCGREGOR C, LIU S, LUAN F S, GAO M L, WENG Y Q. Genetic architecture of fruit size and shape variation in cucurbits: A comparative perspective. Theoretical and Applied Genetics, 2020, 133(1):1-21. doi: 10.1007/s00122-019-03481-3
[31]	MONFORTE A J, DÍAZ A, CAÑO-DELGADO A, KNAAP E. The genetic basis of fruit morphology in horticultural crops: Lessons from tomato and melon. Journal of Experimental Botany, 2014, 65(16):4625-4637. doi: 10.1093/jxb/eru017
[32]	范文林, 王贤磊, 李群, 俞志杰, 李冠. 甜瓜果长基因fl与性别表达基因a的遗传分析及定位. 新疆农业科学, 2018(10):1765-1774.
	FAN W L, WANG X L, LI Q, YU Z J, LI G. Genetic analysis and primary localization of fruit length gene fl and sex expression gene a in melon(Cucumis melo L.). Xinjiang Agricultural Sciences, 2018(10):1765-1774. (in Chinese)

世代 Generation	单株数 Plant number	果面沟表型分离比率 Ratio of fruit sutures	期望比率 Exception ratio	卡方值 χ²	果皮花纹分离比率 Ratio of pericarp pattern	期望比率 Exception ratio	卡方值 χ²
P₁	30	30（有 With）	—	—	0（无 Without）	—	—
P₂	30	0（无 Without）	—	—	30（有 With）	—	—
F₁	30	30（全有 All）	—	—	30（有 With）	—	—
F₂	242	145﹕50 （有﹕无With﹕Without）	3﹕1	0.015	152﹕43 （有﹕无With﹕Without）	3﹕1	0.381
BC₁P₁	150	30（有 With）	—	—	30（有 With）	—	—
BC₁P₂	150	76﹕74 （有﹕无With﹕Without）	1﹕1	0.131	81﹕69 （有﹕无With﹕Without）	1﹕1	0.488

样品名称 Sample ID	序列数 Total reads	碱基数 Total base	测序质量≥30 的碱基数所占百分数 Q30 (%)	G、C 所占总碱基的百分数 GC (%)
1244	38911978	11653958274	92.31	37.01
M5	31878110	9548013962	92.67	36.72
F₂ 群体 F₂ population	2692579	538400027	95.81	36.88
对照 Control	1188503	237656096	96.33	40.87
总量 Total	380446747	83117975446	93.59	36.87

样本 Sample	比对序列数占比 Mapped (%)	测序序列比对参考基因组占比 Properly_mapped (%)	SNP数量 SNP Number	杂合SNP数量 Number of Heter-SNP	纯合SNP数量 Number of Homo-SNP	杂合率 Heter ratio (%)
1244	97.39	90.44	2 250 862	282 705	1 968 157	12.55
MS5	96.56	89.08	1 489 767	146 401	1 343 366	9.82
F₂	97.97	92.50	806 865	60 289	211 029	22.19

染色体 LG	标记数 Number of markers	Gap<5 cM比值 Gap<5 cM ration (%)	秩相关系数 Spearman	最大Gap Max Gap (cM)	总图距 Total distance (cM)	平均图距 Average distance (cM)
1	1267	100.00	0.987	1.74	110.2	0.09
2	504	99.80	0.994	9.33	98.74	0.2
3	1363	100.00	0.989	4.97	111.19	0.08
4	901	100.00	0.995	1.83	138.63	0.15
5	875	100.00	0.992	2.18	94.2	0.11
6	1070	100.00	0.963	3.57	125.08	0.12
7	300	99.00	0.995	6.84	112.25	0.37
8	1104	100.00	0.984	1.83	127.49	0.12
9	649	99.69	0.996	7.98	96.19	0.15
10	591	100.00	0.965	3.05	135.64	0.23
11	1579	100.00	0.982	2.17	137.24	0.09
12	393	99.74	0.996	10.75	97.03	0.25
共计 Total	10596	99.85	0.986	10.75	1383.88	0.13

性状 Traits	QTL位点 QTL position	LOD值LOD threshold	染色体Group ID	起始位点 Start (cM）	基因组位置 Position	结束位点 End (cM)	基因组位置Position	峰值 Max LOD	加性效应ADD	显性效应DOM	贡献率 PVE (%)	候选基因数Candidate gene number
果型指数 Fruit shape (fs)	fs9.1	3.0	9	Marker 1705671 (76.19)	23034060	Marker 1705915 (79.23)	23259152	5.91	0.19	0.05	7.6	23
单果重 Single fruit weight (sfw)	sfw1.1	3.0	1	Marker 297654 (91.06)	34295745	Marker 298408 (91.49)	34435727	3.3	-28.38	-22.21	8.3	17
	sfw2.1	3.0	2	Marker 316077 (18.35)	1626517	Marker 316123 (18.35)	1635504	3.0	17.81	-8.88	3.1	1
	sfw2.2	3.0	2	Marker 414271 (60.38)	18344184	Marker 413648 (60.82)	18881371	5.6	-45.10	-13.59	17.0	15
	sfw6.1	3.0	6	Marker 1186096 (74.87)	30841187	Marker 1186721 (74.89)	38291645	4.2	-21.93	34.30	10.2	354
	sfw7.1	3.0	7	Marker 1254170 (34.54)	4758046	Marker 1255749 (34.97)	7547788	3.2	-25.56	8.55	5.9	113
	sfw10.1	3.0	10	Marker 1718615 (3.05)	485913	Marker 1718692 (3.48)	504601	3.1	-30.64	-14.42	8.3	2
果面沟 Fruit sutures (fst)	fst	—	11	Marker 1993423 (62.18)	20408304	Marker 1998820 (63.05)	21129670	18.3	-0.42	-0.284		33
果皮花纹 Fuit pericarp pattern (fpp)	fpp	—	2	Marker 459584 (90.91)	26112655	Marker 459446 (90.91)	26189137	7.7	0.309	0.087		5
果皮底色 Fruit pericarp color (pc)	fpc	3.0	7	Marker1229174 (7.14)	709451	Marker1229973 (7.14)	892244	4.7	-0.444	-0.09	9.9	15