Images of sections of tissue taken from the upper side of leaning stems of Populus deltoides were projected on a paper grid system. Specimens were oriented so that the cambium and the last three annual increments were included in cross section. Counts of stimulated and non-stimulated cells in randomly selected grids were recorded. This procedure provides a means of subjecting the incidence of tension wood and other cytohistological phenomena to statistical analysis

Berlyn, Graeme P.

English

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A Biometric Technique for Reaction Tissue Research

University of Northern Iowa

Proceedings of the Iowa Academy of ScienceVolume 66 | Annual Issue Article 121959A Biometric Technique for Reaction TissueResearchGraeme P. BerlynIowa State UniversityCopyright © Copyright 1959 by the Iowa Academy of Science, Inc.Follow this and additional works at: https://scholarworks.uni.edu/piasThis Research is brought to you for free and open access by UNI ScholarWorks. It has been accepted for inclusion in Proceedings of the Iowa Academyof Science by an authorized editor of UNI ScholarWorks. For more information, please contact scholarworks@uni.edu.Recommended CitationBerlyn, Graeme P. (1959) "A Biometric Technique for Reaction Tissue Research," Proceedings of the Iowa Academy of Science: Vol. 66:No. 1 , Article 12.Available at: https://scholarworks.uni.edu/pias/vol66/iss1/12A Biometric Technique for Reaction Tissue Hesearch 1 By GRAEME P. BERLYN Abstract. Images of sections of tissue taken from the upper side of leaning stems of Populus deltoides were projected on a paper grid system. Specimens were oriented so that the cambium and the last three annual increments were included in cross section. Counts of stimulated and non-stimulated cells in randomly selected grids were recorded. This procedure provides a means of subjecting the incidence of tension wood and other cytohistological phe-nomena to statistical analysis. The measurement of specific cellular characteristics can be of fun-damental value in the interpretation of cytological and histological phenomena such as the formation of the so-called tension wood of various angiosperms and compression wood of gymnosperms. The term reaction tissue is used to refer to these tissues which are found in leaning (or otherwise stimulated) stems and branches. The re-action tissue of angiosperms commonly occurs on the upper side of leaning stems, and the woody tissue from this region is both phys-ically and chemically different from non-reaction tissue (Wardrop and Dadswell, 1948). During the study of the reaction tissue of eastern cottonwood (Populus deltoides Bartr.), it became necessary to develop a system of micrometry that would be consistent with modern statistical com-putation and analysis. Recent developments in both computers and statistical theory open many new possibilities in biological research, and valuable information can be gained from otherwise obscure sources. These techniques can be valuable in phylogenetic and com-parative studies as well as experimental botanical research (cf. Bailey, 1953). MICROTECHNIQUE Stem material was collected at breast height from the upper side of leaning cottonwood trees. Cores of tissue were extracted with a modified 0 inch plug cutter used in an ordinary brace and bit. Sec-tions were cut on the sliding microtome without embedding. The specimens were oriented so that the cambium and the last three 1 Journal Paper l\"o. J-3665 of the Iowa Agricultural and Home Economics Experiment Station, Ames, Iowa. Project No. 1330, conducted in cooperation with the Ames Branch of the Central States Forest Experiment Station, Ames, Iowa. Sincere appreciation is due Dr. D. W. Bensend, Dr. J. E. Sass, and the Forest Products -Laboratory, Madison, Wisconsin, for advice and encouragement during the course of this study. The cooperation of Dr. H. T. David and P. G.- Homeyer of the Iowa State College Statistical Laboratory is gratefully acknowledged. 98 1Berlyn: A Biometric Technique for Reaction Tissue ResearchPublished by UNI ScholarWorks, 19591959] REACTION TISSUE 99 annual increments were included in cross section on the sections. The sections were then stained in chloroiodide of zinc. This tem-porary stain is specific for the cellulosic material of the so-called gelatinous layer of the cell wall of tension wood fibers. This gelat-inous layer of the stimulated cells stains dark blue, and in eastern cottonwood frequently swells into the cell lumen in response to this stain (cf. Wahlgren, 1956). Coverslips were sealed with wax. TREE No.: __ TENSION WOOD RESEARCH PROJECT OTHER: PLOT No.:__ S. I_ LEAN:_0 CORE: I_ 2_3_ RING No.1_2_3_ RING WIDTH·._ FIELD No. __ I: No. CELLS:_ l: GEL FBRS: _ 0 I 2 3 4 56 7 8 9 ,-----.--------.---- - - ,.......--,------~---0 I 2 3 4 f---+--1---L-- --~-+--f---f----!---5 6 7 ' 8 I 1--~1---9: 1. _L_~HJ I _ _J Figure 1. Sample grid shC'et. MrcROMETRY The slides were projected vertically at magnification 139 X with a microprojector using a ribbon filament lamp. Carbon arcs supply too much heat for these temporary mounts and are therefore un-suitable. The image of the tissue with its three included annual increments was projected on a paper grid system (Figure 1) which was taped to the desk surface. Observations for each microscopic field were recorded on separate grid sheets. These sheets serve as a permanent' record. The multilithed 10 by 10 grid measures 12 .5 cm by 12 .5 cm. Each grid square is a 1.2 5 cm square. At 139 X the entire grid system covers an area of tissue 0.9 mm by 0.9 mm or 2Proceedings of the Iowa Academy of Science, Vol. 66 [1959], No. 1, Art. 12https://scholarworks.uni.edu/pias/vol66/iss1/12100 IOWA ACADEMY OF SCIENCE [Vol. 66 810,000 square microns. Each of the 100 individual grid squares then encloses 0.09 mm by 0.09 mm of tissue or 8100 square microns. In each of the individual grid squares selected for observation, actual counts were made of stimulated and nonstimulated cells. The data were recorded directly in the individual grid squares. In the binomial system used, the first number in a particular grid square refers to the number of cells with a gelatinous layer and the second number refers to nonstimulated cells. It is generally not necessary or practical to take data for every square of the grid system. In this study 8 grid squares were ran-domly selected for observation on each grid system. This was con-veniently done with the aid of a random number table (Snedecor, 19 5 6). Three grid systems were analyzed for each of three annual increments, making a total of 9 grid systems per slide. Since 8 grid squares were analyzed per grid system, data were recorded for 72 grid squares. If each grid square contained about 20 cells there would be data on 1,440 cells for every slide. For more detailed analysis 10 to 15 grid squares per grid system could be analyzed without excessively increasing the time of analysis. Data taken in this manner fit nicely into a wide variety of statistical analyses. Table 1 Data from Plot 5 site index 97 Lean Proportion Gelatinous (degrees) Ring Width fibers in sample Tree No. Year x (mm.) y 31 1956 3 2.14 0.000 1955 3 1.75 0.000 1954 3 1.54 0.000 30 1956 5 0.60 0.000 1955 5 1.00 0.000 1954 5 1.09 0.000 34 1956 6 2.96 0.201 1955 6 2.36 0.0196 1954 6 2.90 0.0775 29 1956 9 1.42 0.341 1955 9 1.25 0.291 1954 9 1.57 0.390 33 1956 12 2.33 0.945 1955 12 1.80 0.860 1954 12 1.21 0.591 32 1956 15 2.57 0.786 1955 15 0.96 0.795 1954 15 1.14 0.876 Regression analysis of Y on X : i!"=0.343 + 0.8114 (X-X) r=0.946** 3Berlyn: A Biometric Technique for Reaction Tissue ResearchPublished by UNI ScholarWorks, 19591959] REACTION TISSUE 101 RESULTS The foregoing method is being used in connection with a study on the effect of environmental and genetic factors on the reaction tissue of eastern cottonwood. For purposes of illustration some raw data (Table 1) were taken from a cottonwood grove along the Mis-souri River in western Iowa. A simple linear regression of proportion of gelatinous fibers on lean was calculated. In this particular plot lean accounted for 89.4 per cent of the variation as measured by the sums of squares. The plot of the raw data has not been subjected to any smoothing function and merely represents the data as re-corded (Figure 2). The graph shows three points for each lean class sampled. These three points above a specific lean value repre-sent three annual increments from the same tree. Each point is identified as to year and the corresponding ring width for that year is recorded to the left of the point. (/) (ii <( al a: 1.0 <( LU >-' ~08 LU al LL r.n0.6 ::> 0 ::> z ;::: 0.4 <( ..J LU <!) LL 02 0 z 0 i= a: 0 a. 0 a: a. Figure 2. 2 2.14 17!5. I 09 4 2.96Q 2.9oe ~:ig.2.36(1 1.09 2:330 1.eoct 114e 01956 YEAR'S GROWTH () 1955 YEAR'S GROWTH &1954 YEAR'S GROWTH e ALL 3 YEAR'S Number to left of point is ring width in mm. 6 8 10 12 14 16 LEAN - BREAST HEIGHT -COTTONWOOD 18 Sample data from a stand of eastern cottonwood showing relationships between lean ring width, year, and proportion of gelatinous fibers. SUMMARY The procedures described herein provide a means of subjecting the incidence of tension wood and other cytohistological phenomena to statistical analysis in relation to various environmental and genetic factors. 4Proceedings of the Iowa Academy of Science, Vol. 66 [1959], No. 1, Art. 12https://scholarworks.uni.edu/pias/vol66/iss1/12102 IOWA ACADEMY OF SCIENCE [Vol. 66 Literature Cited Bailey, I. W. 1953. Evolution of the tracheary tissue of land plants. Amer. J our. Bot. 40: 4-8. Snedecor, George W. 1956. Statistical methods. 5th edition. Iowa State College Press, Ames, Iowa, pp. 10-13. Wahlgren, Harold E. 1956. Effect of tension wood on the longitudinal shrink-age and specific gravity of eastern cottonwood. M.S. Thesis, Library, Iowa State College, pp. 56. Wardrop, A. B. and Dadswell, H. E. 1948. The nature of reactionwood. I. The structure and properties of tension wood fibers. Australian J our. of Scientific Research, Series B, Biological Sciences, 1 :3-16. DEPARTMENT OF FORESTRY lowA STATE UNIVERSITY AMES, lowA 5Berlyn: A Biometric Technique for Reaction Tissue ResearchPublished by UNI ScholarWorks, 1959

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A Biometric Technique for Reaction Tissue Research

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