Chin J Plan Ecolo ›› 2018, Vol. 42 ›› Issue (9): 917-925.doi: 10.17521/cjpe.2018.0087

• Research Articles • Previous Articles     Next Articles

What is the optimal number of leaves when measuring leaf area of tree species in a forest community?

GAO Si-Han,GE Yu-Xi,ZHOU Li-Yi,ZHU Bao-Lin,GE Xing-Yu,LI Kai(),NI Jian()   

  1. College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua, Zhejiang 321004, China
  • Received:2018-04-16 Revised:2018-06-09 Online:2018-06-11 Published:2018-09-20
  • Contact: Kai LI,Jian NI;
  • Supported by:
    Supported by the National Natural Science Foundation of China(41471049)


Aims Leaf size, as one of the easier measured plant morphological traits, reflects response and adaptation of plants to environment and indicates functions and processes of ecosystem. When measuring leaf size (the leaf area, LA) on the field, the common accepted practice considers that the number of leaves picked off is often 10-20. However, what is the optimal number of leaves remains unknown. In this study, we attempt to determine how many leaves should be investigated when the leaf size of a tree is measured.

Methods This study selected two dominant tree species (Schima superba, Ss and Liquidambar formosana, Lf) from a broadleaved evergreen and deciduous mixed forest in Jinhua Mountain of Zhejiang Province, eastern China. On the basis of sampling (>2 500 leaves for each tree) in five classes of the diameter of breast height (DBH) of tree species and at six directions for each individual, variations of LA in the two tree species are statistically analyzed. The optimal number of leaves, which can mostly represent the common leaf size feature, is further investigated.

Important findings Mean LAof the evergreen tree Ss was smaller than that of the deciduous tree Lf. The former was (41.60 ± 10.88) cm 2 (16.74-100.80 cm 2) and the latter was (57.65 ± 19.35) cm 2 (11.31-129.51 cm 2). LA of Lf was significantly related to the DBH, but LAof Ss was not. LA of both trees in the middle DBH class (15-20 cm) was not significantly different from their means. LA of two trees have no significant correlations with the sampling directions, but LA at the east, west and bottom had no significant relationships with their means. Considering the representativeness and practicality in the field sampling, the priority of selecting leaves can target the bottom direction of middle diameter mature trees. Random sampling analysis indicated that, the optimal number of leaves for tree LA measurement is species specific. The optimal number of leaves for Ss is 40 and for Lf is at least 170, respectively. Therefore, when measuring leaf area in a forest community, the optimal sampling number of leaves should not be limited to 10-20 leaves. Under sufficient labor, material and time, more leaves should be measured.

Key words: plant functional traits, leaf area, sampling number, sampling direction, tree age, evergreen and deciduous broadleaved mixed forest

Fig. 1

Relationship between leaf area and diameter at breast height of two tree species. A, B, Quartile map; C, D, Scatter plot; E, F, Confidence interval. Ss, Lf, mean values of all samples in all classes of diameter at breast height for Schima superba and Liquidambar formosana, respectively. D2-D6, D2-D6 classes of diameter at breast height. *, p < 0.05; **, p < 0.01."

Fig. 2

Relationship between leaf area and sampling direction of two tree species. A, B, Quartile map; C, D, Scatter plot; E, F, Confidence interval. *, p < 0.05; **, p < 0.01."

Fig. 3

Deviation of leaf area to diameter and sampling direction of two tree species. A, Diameter of Ss; B, Sampling direction of Ss; C, Diameter of Lf; D, Sampling direction of Lf. *, p < 0.05; **, p < 0.01. The vertical line indicates the mean value returning to zero."

Fig. 4

Deviation of mature and non-mature leaf areas to diameter at breast height (DBH) of two tree species. A, Schima superba (Ss). B, Liquidambar formosana (Lf). The horizontal line indicates the mean value returning to zero."

Fig. 5

Mean of random sampling, 95% confidence interval and significant test of leaf areas of two tree species. A, C, All leaf samples. B, D, Leaf samples of medium diameter class and bottom sampling direction. The black line of the leaf area plot indicates the mean value of random sampling, and the red line indicates the 95% confidence interval; the light blue block is the area of significant difference, indicating that the sample size of the right position of the block is the optimal sampling number of leaves. Ss, Schima superba; Lf, Liquidambar formosana."

[1] Aerts R ( 1995). The advantages of being evergreen. Trends in Ecology and Evolution, 10, 402-407.
doi: 10.1016/S0169-5347(00)89156-9 pmid: 21237084
[2] Albert CH, Thuiller W, Yoccoz NG, Soudant A, Boucher F, Saccone P, Lavorel S ( 2010). Intraspecific functional variability: Extent, structure and sources of variation. Journal of Ecology, 98, 604-613.
doi: 10.1111/j.1365-2745.2010.01651.x
[3] Auger S, Shipley B ( 2013). Inter-specific and intra-specific trait variation along short environmental gradients in an old-growth temperate forest. Journal of Vegetation Science, 24, 419-428.
doi: 10.1111/jvs.2013.24.issue-3
[4] Bolnick DI, Amarasekare P, Araojo MS, Burger R, Levine JM, Novak M, Rudolf VHW, Schreiber SJ, Urban MC, Vasseur DA ( 2011). Why intraspecific trait variation matters in community ecology. Trends in Ecology and Evolution, 26, 183-192.
doi: 10.1016/j.tree.2011.01.009 pmid: 21367482
[5] Bu WS, Schmid B, Liu XJ, Li Y, Hardtle W, von Oheimb G, Liang Y, Sun ZK, Huang YY, Bruelheide H, Ma KP ( 2017). Interspecific and intraspecific variation in specific root length drives aboveground biodiversity effects in young experimental forest stands. Journal of Plant Ecology, 10, 158-169.
doi: 10.1093/jpe/rtw096
[6] Chen YT, Xu ZZ ( 2014). Review on research of leaf economics spectrum. Chinese Journal of Plant Ecology, 38, 1135-1153.
doi: 10.3724/SP.J.1258.2014.00108
[ 陈莹婷, 许振柱 ( 2014). 植物叶经济谱的研究进展. 植物生态学报, 38, 1135-1153.]
doi: 10.3724/SP.J.1258.2014.00108
[7] Cornelissen JHC, Lavorel S, Garnier E, Diaz S, Buchmann N, Gurvich DE, Reich PB, Ter Steege H, Morgan HD, van der Heijden MGA, Pausas JG, Poorter H ( 2003). A handbook of protocols for standardized and easy measurement of plant functional traits worldwide. Australian Journal of Botany, 51, 335-380.
doi: 10.1071/BT02124
[8] Díaz S, Kattge J, Cornelissen JHC, Wright IJ, Lavorel S, Dray S, Reu B, Kleyer M, Wirth C, Prentice IC, Garnier E, Boenisch G, Westoby M, Poorter H, Reich PB, Moles AT, Dickie J, Gillison AN, Zanne AE, Chave J, Wright SJ, Sheremet’ev S, Jactel H, Baraloto C, Cerabolini B, Pierce S, Shipley B, Kirkup D, Casanoves F, Joswig J, Günther A, Falczuk V, Rüger N, Mahecha MD, Gorné LD ( 2016). The global spectrum of plant form and function. Nature, 529, 167-171.
doi: 10.1038/nature16489 pmid: 26700811
[9] Donovan LA, Maherali H, Caruso CM, Huber H, de Kroon H ( 2011). The evolution of the worldwide leaf economics spectrum. Trends in Ecology and Evolution, 26, 88-95.
doi: 10.1016/j.tree.2010.11.011 pmid: 21196061
[10] Fang F, Guo SL ( 2004). DCA ordination on woody plant communities in Beishan Mountain of Jinhua, Zhejiang Province. Journal of Zhejiang Normal University (Natural Science), 27(1), 55-61.
doi: 10.3969/j.issn.1001-5051.2004.01.014
[ 方芳, 郭水良 ( 2004). 浙江金华北山木本植物群落的DCA排序. 浙江师范大学学报(自然科学版), 27(1), 55-61.]
doi: 10.3969/j.issn.1001-5051.2004.01.014
[11] Givnish TJ ( 2002). Adaptive significance of evergreen vs. deciduous leaves: Solving the triple paradox. Silva Fennica, 36, 703-743.
[12] Guo SL, Liu P, Chen G, Lu X ( 1993). Observations on the floristic and vegetation in Beishan Mountain of Jinhua in Zhejiang Province. Journal of Zhejiang Normal University (Natural Science), 16(2), 59-67.
[ 郭水良, 刘鹏, 陈刚, 卢晓 ( 1993). 浙江金华北山植物区系及植被. 浙江师范大学学报(自然科学版), 16(2), 59-67.]
[13] Li MC, Zhu JJ, Sun YR ( 2009). Responses of specific leaf area of dominant tree species in Northeast China secondary forests to light intensity. Chinese Journal of Ecology, 28, 1437-1442.
[ 李明财, 朱教君, 孙一荣 ( 2009). 东北次生林主要树种比叶面积对光照强度的响应. 生态学杂志, 28, 1437-1442.]
[14] Liu XJ, Ma KP ( 2015). Plant functional traits—Concepts, applications and future directions. Scientia Sinica Vitae, 45, 325-339.
doi: 10.1360/N052014-00244
[ 刘晓娟, 马克平 ( 2015). 植物功能性状研究进展. 中国科学: 生命科学, 45, 325-339.]
doi: 10.1360/N052014-00244
[15] McDonald PG, Fonseca CR, Overton J, Westoby M ( 2003). Leaf-size divergence along rainfall and soil-nutrient gradients: Is the method of size reduction common among clades? Functional Ecology, 17, 50-57.
doi: 10.2307/3599027
[16] Meng TT, Ni J, Wang GH ( 2007). Plant functional traits, environments and ecosystem functioning. Journal of Plant Ecology (Chinese Version), 31, 150-165.
doi: 10.17521/cjpe.2007.0019
[ 孟婷婷, 倪健, 王国宏 ( 2007). 植物功能性状与环境和生态系统功能. 植物生态学报, 31, 150-165.]
doi: 10.17521/cjpe.2007.0019
[17] Messier J, Lechowicz MJ, McGill BJ, Violle C, Enquist BJ ( 2017). Interspecific integration of trait dimensions at local scales: The plant phenotype as an integrated network. Journal of Ecology, 105, 1775-1790.
doi: 10.1111/1365-2745.12755
[18] Peppe DJ, Royer DL, Cariglino B, Oliver SY, Newman S, Leight E, Enikolopov G, Fernandez-Burgos M, Herrera F, Adams JM, Correa E, Currano ED, Erickson JM, Hinojosa LF, Hoganson JW, Iglesias A, Jaramillo CA, Johnson KR, Jordan GJ, Kraft NJ, Lovelock EC, Lusk CH, Niinemets U, Pe?uelas J, Rapson G, Wing SL, Wright IJ ( 2011). Sensitivity of leaf size and shape to climate: Global patterns and paleoclimatic applications. New Phytologist, 190, 724-739.
doi: 10.1111/j.1469-8137.2010.03615.x pmid: 21294735
[19] Pérez-Harguindeguy N, Diaz S, Garnier E, Lavorel S, Poorter H, Jaureguiberry P, Bret-Harte MS, Cornwell WK, Craine JM, Gurvich DE, Urcelay C, Veneklaas EJ, Reich PB, Poorter L, Wright IJ, Ray P, Enrico L, Pausas JG, de Vos AC, Buchmann N, Funes G, Quétier F, Hodgson JG, Thompson K, Morgan HD, ter Steege H, van der Heijden MG A, Sack L, Blonder B, Poschlod P, Vaieretti MV, Conti G, Staver AC, Aquino S, Cornelissen JHC ( 2013). New handbook for standardised measurement of plant functional traits worldwide. Australian Journal of Botany, 61, 167-234.
doi: 10.1071/BT12225
[20] Royer DL, McElwain JC, Adams JM, Wilf P ( 2008). Sensitivity of leaf size and shape to climate within Acer rubrum and Quercus kelloggii. New Phytologist, 179, 808-817.
[21] Siefert A, Violle C, Chalmandrier L, Albert CH, Taudiere A, Fajardo A, Aarssen LW, Baraloto C, Carlucci MB, Cianciaruso MV, Dantas VD, DeBello F, Duarte LDS, Fonseca CR, Freschet GT, Gaucherand S, Gross N, Hikosaka K, Jackson B, Jung V, Kamiyama C, Katabuchui M, Kembel SW, Kichenin E, Kraft NJB, Lagerstrom A, Le Bagousse-Pinguer Y, Li YZ, Mason N, Messier J, Nakashizuka T, Overton JM, Peltzer DA, Perez-Ramos IM, Pillar VD, Prentice HC, Richardson S, Sasaki T, Schamp BS, Vandewalle M, Wardle DA ( 2015). A global meta-analysis of the relative extent of intraspecific trait variation in plant communities. Ecology Letters, 18, 1406-1419.
doi: 10.1111/ele.12508 pmid: 26415616
[22] Violle C, Enquist BJ, McGill BJ, Jiang L, Albert CH, Huishof C, Jung V, Messier J ( 2012). The return of the variance: Intraspecific variability in community ecology. Trends in Ecology and Evolution, 27, 244-252.
doi: 10.1016/j.tree.2011.11.014
[23] Wang CS, Wang SP ( 2015). A review of research on responses of leaf traits to climate change. Chinese Journal of Plant Ecology, 39, 206-216.
doi: 10.17521/cjpe.2015.0020
[ 王常顺, 汪诗平 ( 2015). 植物叶片性状对气候变化的响应研究进展. 植物生态学报, 39, 206-216.]
doi: 10.17521/cjpe.2015.0020
[24] Westoby M, Falster DS, Moles AT, Vesk PA, Wright IJ ( 2002). Plant ecological strategies: Some leading dimensions of variation between species. Annual Review of Ecology and Systematics, 33, 125-159.
doi: 10.1146/annurev.ecolsys.33.010802.150452
[25] Wright IJ, Reich PB, Westoby M, Ackerly DD, Baruch Z, Bongers F, Cavender-Bares J, Chapin T, Cornelissen JHC, Diemer M, Flexas J, Garnier E, Groom PK, Gulias J, Hikosaka K, Lamont BB, Lee T, Lee W, Lusk C, Midgley JJ, Navas ML, Niinemets U, Oleksyn J, Osada N, Poorter H, Poot P, Prior L, Pyankov VI, Roumet C, Thomas SC, Tjoelker MG, Veneklaas EJ, Villar R ( 2004). The worldwide leaf economics spectrum. Nature, 428, 821-827.
doi: 10.1038/nature02403
[1] Shitong Wang Yaozhan Xu Teng Yang Xinzeng Wei Mingxi Jiang. Impacts of microhabitats on leaf functional traits characteristics of Sinojackia huangmeiensis [J]. Biodiv Sci, 2020, 28(3): 0-0.
[2] LI Qun, ZHAO Cheng-Zhang, WANG Ji-Wei, WEN Jun, LI Zi-Qin, MA Jun-Yi. Morphological and photosynthetic physiological characteristics of Saussurea salsa in response to flooding in salt marshes of Xiao Sugan Lake, Gansu, China [J]. Chin J Plant Ecol, 2019, 43(8): 685-696.
[3] ZHAO Dan-Dan, MA Hong-Yuan, LI Yang, WEI Ji-Ping, WANG Zhi-Chun. Effects of water and nutrient additions on functional traits and aboveground biomass of Leymus chinensis [J]. Chin J Plant Ecol, 2019, 43(6): 501-511.
[4] WANG Jin, ZHU Jiang, AI Xun-Ru, YAO Lan, HUANG Xiao, WU Man-Ling, ZHU Qiang, HONG Jian- Feng. Effects of topography on leaf functional traits across plant life forms in Xingdou Mountain, Hubei, China [J]. Chin J Plant Ecol, 2019, 43(5): 447-457.
[5] YANG Huan-Ying, SONG Jian-Da, ZHOU Tao, JIN Guang-Ze, JIANG Feng, LIU Zhi-Li. Influences of stand, soil and space factors on spatial heterogeneity of leaf area index in a spruce-fir valley forest in Xiao Hinggan Ling, China [J]. Chin J Plant Ecol, 2019, 43(4): 342-351.
[6] Liu Qiang, Cai Erli, Zhang Jialin, Song Qiao, Li Xiuhong, Dou Baocheng. A Modification of the Finite-length Averaging Method in Measuring Leaf Area Index in Field [J]. Chin Bull Bot, 2018, 53(5): 671-685.
[7] Guo Shulei, Zhang Jun, Qi Jianshuang, Yue Runqing, Han Xiaohua, Yan Shufeng, Lu Caixia, Fu Xiaolei, Chen Nana, Ku Lixia, Tie Shuanggui. Analysis of Meta-quantitative Trait Loci and Their Candidate Genes Related to Leaf Shape in Maize [J]. Chin Bull Bot, 2018, 53(4): 487-501.
[8] PENG Xi, YAN Wen-De, WANG Feng-Qi, WANG Guang-Jun, YU Fang-Yong, ZHAO Mei-Fang. Specific leaf area estimation model building based on leaf dry matter content of Cunninghamia lanceolata [J]. Chin J Plan Ecolo, 2018, 42(2): 209-219.
[9] Qun LI, Cheng-Zhang ZHAO, Lian-Chun ZHAO, Jian-Liang WANG, Wei-Tao ZHANG, Wen-Xiu YAO. Empirical relationship between specific leaf area and thermal dissipation of Phragmites australis in salt marshes of Qinwangchuan [J]. Chin J Plan Ecolo, 2017, 41(9): 985-994.
[10] Ze-Bin LIU, Yan-Hui WANG, Yu LIU, Ao TIAN, Ya-Rui WANG, Hai-Jun ZUO. Spatiotemporal variation and scale effect of canopy leaf area index of larch plantation on a slope of the semi-humid Liupan Mountains, Ningxia, China [J]. Chin J Plan Ecolo, 2017, 41(7): 749-760.
[11] GAO Lin, WANG Xiao-Fei, GU Xing-Fa, TIAN Qing-Jiu, JIAO Jun-Nan, WANG Pei-Yan, LI Dan. Exploring the influence of soil types underneath the canopy in winter wheat leaf area index remote estimating [J]. Chin J Plan Ecolo, 2017, 41(12): 1273-1288.
[12] Jia-Xiang LI, Wen-Ting XU, Gao-Ming XIONG, Yang WANG, Chang-Ming ZHAO, Zhi-Jun LU, Yue-Lin LI, Zong-Qiang XIE. Leaf nitrogen and phosphorus concentration and the empirical regulations in dominant woody plants of shrublands across southern China [J]. Chin J Plan Ecolo, 2017, 41(1): 31-42.
[13] Ling HAN, Cheng-Zhang ZHAO, Ting XU, Wei FENG, Bei-Bei DUAN, Hui-Ling ZHENG. Trade-off between leaf size and vein density of Achnatherum splendens in Zhangye wetland [J]. Chin J Plan Ecolo, 2016, 40(8): 788-797.
[14] Jing GAO, Jin-Niu WANG, Bo XU, Yu XIE, Jun-Dong HE, Yan WU. Plant leaf traits, height and biomass partitioning in typical ephemerals under different levels of snow cover thickness in an alpine meadow [J]. Chin J Plan Ecolo, 2016, 40(8): 775-787.
[15] Ming ZHOU, Zhi-Li LIU, Guang-Ze JIN. Improving the accuracy of indirect methods in estimating leaf area index using three correction schemes in a Larix gmelinii plantation [J]. Chin J Plan Ecolo, 2016, 40(6): 574-584.
Full text



[1] Lin liang-qiu;Zhang Qing-qi and Wu Wen-shan. A Study on Pollen Morphology of Rosa laevigata and Its Nutrition Composition[J]. Chin Bull Bot, 1994, 11(04): 43 -44 .
[2] Jie Dong;Fenghui Qi;Yaguang Zhan. Establishment of the Suspension Culture System and Optimization of Biosynthesis of Gallic Acid in Acer ginnala[J]. Chin Bull Bot, 2008, 25(06): 734 -740 .
[3] Li Guo-zhen;Qin Ming-bo;Kang Ning-ling;Xie De-yu;Ye He-chun and Li Guo-feng. Tissue Culture and Chromosome Analysis of Arnebia euchroma[J]. Chin Bull Bot, 1992, 9(01): 37 -41 .
[4] Han Bi-wen. The Synthitic Activities of Roots and their Relation to the Above-grond Parts[J]. Chin Bull Bot, 1984, 2(23): 23 -25 .
[5] Guiling Wang;Zhiwei Qin;Xiuyan Zhou;Zhiyun Zhao. Genetic Analysis and SSR Markers of Tuberculate Trait in Cucumis sativus[J]. Chin Bull Bot, 2007, 24(02): 168 -172 .
[6] Hui Yang;Lizhe An;Zhiye Wang;Jianping Zhou;Xunling Wang. Effects of Enhanced UV-B Radiation on Pollen Activities of 2 Tomato Cultivars in Terms of Endogenous Hormone,Polyamine and Proline Levels in Stamens[J]. Chin Bull Bot, 2007, 24(02): 161 -167 .
[7] Hao Zhao;Zhao Xue-chen;Zheng Shu-jun and Qu Chun-ying. Winter Hardiness of Wheat Seedling at Differant Leaf-Age[J]. Chin Bull Bot, 1985, 3(05): 38 -40 .
[8] Chang Huey-ju;Guan Zhong-tian;Zhou Lin and Hsu Kuo-shih. Comparison of two natural cycad communities in China[J]. Chin Bull Bot, 1995, 12(专辑): 52 -58 .
[9] . Positional Information and Plant Development [J]. Chin Bull Bot, 2005, 22(03): 366 -374 .
[10] Deyong Ren, Guanghua He, Yinghua Ling, Xianchun Sang, Zhenglin Yang, Fangming Zhao. Analysis of Quantitative Trait Loci Additive and Epistasis Effects for Panicle Length with Single Segment Substitution Lines in Rice[J]. Chin Bull Bot, 2010, 45(06): 662 -669 .