Measuring Roots exploring the hidden half. Jhonathan E. Ephrath
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1 Measuring Roots exploring the hidden half Jhonathan E. Ephrath I-Core course -Plant Functional
2 Plant activity and the distribution of photosynthates between the root and the shoot will determine the development of the root system: 1. Total length of the root system 2. Root depth I-Core course -Plant Functional
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4 Plant activity and the distribution of photosynthates between the root and the shoot will determine the development of the root system: 1. Total length of the root system 2. Root depth 3. Root branching and root lateral root length 4. Root diameter 5. Root volume and root surface area I-Core course -Plant Functional
5 6. The position and the appearance of root hairs in relation to root tip 7. Total length of root hairs. I-Core course -Plant Functional
6 Since most of the agricultural crop are annuals, the elongation of the root system start right from the root that develop from the seed. In Monocotyledon, right after this initial development, the development of new root will develop from higher nodes on the stem, so the root system is buikt of many development axes and their branching. In dicotyledons, a tap root will be developed with many lateral roots and their branching. Another option is a diffuse root system with lateral roots that will be function as tap root.
7 Vertical axis A Lateral root of 1 st order B A B Lateral root of higher order C C
8 In the figure we can see 3 types of root systems: A represent the tap root pattern:
9 This type of root contain a main root that grow downward and lateral roots that their main function is transporting. Although the tap roots absorb all kind of materials while they are young, most of their life, these roots transport water and nutrients absorbed by the lateral roots to the shoot and to deliver photosynthates towards the young roots and to the growth meristems.
10 Type B roots, and lateral roots of FIRST order. They can be horizontal or vertical and they can grow at the begining in an angle and then vertically. The main function of these roots is absorbing and transporting, depending on the crop and on the season. A C B
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12 Type C roots are higher order roots and they are mainly ABSORBING roots. These roots can be temporary roots (which might die). A B C
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14 In order to monitor the different aspects of the root system, various methods had been developed throughout the years. I-Core course -Plant Functional
15 Soil cores I-Core course -Plant Functional
16 Using this method, a driller is inserted into the soil and pushed out with soil sample. The driller can be of different length, depending on the soil type, depth of measurements and the equipment used in order to push the driller into the soil: I-Core course -Plant Functional
17 I-Core course -Plant Functional
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20 I-Core course -Plant Functional
21 I-Core course -Plant Functional Gambia '05, Seminar
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23 Separating the roots from the soil by washing or by air pressure I-Core course -Plant Functional
24 I-Core course -Plant Functional
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26 I-Core course -Plant Functional
27 I-Core course -Plant Functional
28 I-Core course -Plant Functional EC 8 ds/m
29 Experimental set up I-Core course -Plant Functional
30 Shoot length (cm) 150 EC 1.0 ds/m EC 8.0 ds/m 100 * * * * 50 * 0 Date Figure 1. Shoot length of tomato under non-saline & saline water irrigation Values are means of sixteen replicates and standard error bars. Asterix mark indicates significant difference between treatments of the same time interval (P =0.05, Student s t-test) I-Core course -Plant Functional
31 In-growth core measurements In-growth core EC 1.0 ds/m EC 8.0 ds/m I-Core course -Plant Functional
32 Root length density (cm/cm 3 ) Root mass density (mg/cm 3 ) Total root length (cm) Root surface area (cm 2 ) * (a) * (b) * (c) * (d) ds/m 8.0 ds/m 1.0 ds/m 8.0 ds/m Figure 5. Root length (a), root surface area (b), root length density (c) and root mass density (d) of tomato I-Core under course non-saline -Plant Functional & saline water irrigation
33 In growth cores for measuring roots I TAHAL_2018
34 TAHAL_2018
35 Ingrowth core
36 Ingrowth core
37 TAHAL_2018
38 TAHAL_2018
39 I-Core course -Plant Functional
40 Root Length Density (RLD) RLD LENGTH [ cm (roots)] VOLUME [ cm (soil)] 3 I-Core course -Plant Functional
41 Line intersection method for estimating root length Newman (1966): L NA 2H L Root length N No. of intersections A Area of tray H Total length of grids on tray I-Core course -Plant Functional
42 I-Core course -Plant Functional Gambia '05, Seminar
43 L NA 2H d b A (Tray area) = a*b H = length of grid lines H a b b a d d 2ab d a d L N a b * * * ab 2* 2* d L N = I-Core course -Plant Functional 4 d
44 L d dimenssion of square s side N No. of intesections 4 d N If d=1cm, we can write the equation as- L N 4 I-Core course -Plant Functional Tennant (1975)
45 In our example, in case we have a root that has 7 inersctions with a grid, in which the dimenssion of each square is 1X1 cm, d d L 4 N L cm 4 I-Core course -Plant Functional
46 Using this method cause several problems: 1. The amount of labor involved in taking the samples and analyze them 2. More problematic: The information gained by in this method is correct for the time the sample was taken We do not know what happened before we took the measurement, and since this method is destructive, we will not know what will happen later on: This method cannot provide us with the dynamic development of the roots. I-Core course -Plant Functional
47 Trying to solve the problem of studying root dynamic development over a long period of time, the rhizotrons were developped: I-Core course -Plant Functional
48 I-Core course -Plant Functional
49 I-Core course -Plant Functional
50 I-Core course -Plant Functional
51 A view of a root and root hairs as viewd in a rhizotron with a binocular Microscope. (the root hairs are approx. 100 mm). I-Core course -Plant Functional
52 I-Core course -Plant Functional Gambia '05, Seminar
53 I-Core course -Plant Functional
54 I-Core course -Plant Functional
55 The Rhizotron system has also disadvantages: 1. Very expensive facility 2. The soil is disturbed and therefore affects the root development 3. Since the size of the facility is relatively small, the number of treatments and replicates is very limited. Therefore a cheeper version of the rhizotron was debelopped the minirhizotron I-Core course -Plant Functional
56 minirhizotron I-Core course -Plant Functional
57 I-Core course -Plant Functional
58 The minirhizotron system I-Core course -Plant Functional
59 I-Core course -Plant Functional
60 I-Core course -Plant Functional
61 I-Core course -Plant Functional
62 Four light bulbs I-Core course -Plant Functional
63 I-Core course -Plant Functional
64 I-Core course -Plant Functional
65 Indexing handle Camera head I-Core course -Plant Functional
66 Camera head U.V. Light Camera I-Core course -Plant Functional Vis. light
67 Indexing handle Camera head I-Core course -Plant Functional
68 I-Core course -Plant Functional
69 Why use Minirhizotrons? Direct observation of the root segment Non-destructive Repeated onservations Quantify fine root phenology Quantify fine root production, growth and mortality (root turnover) See more than just fine roots: - Soil organisms - mycorhizae Minirhizotrons are improving I-Core course -Plant Functional
70 I-Core course -Plant Functional
71 I-Core course -Plant Functional
72 18 mm 0.3 mm 12.5 mm 0.1 mm 0.5 mm שורשים של כותנה בגילים שונים I-Core course -Plant Functional
73 180 GDD; D 20 cm 300 GDD; D 20 cm 240 GDD; D 20 cm 360 GDD; D 20 cm 18mm I-Core course -Plant Functional
74 Roots and mycorrhizae I-Core course -Plant Functional
75 Experimental set up I-Core course -Plant Functional
76 Minirhizotron measurements Minirhizotron system EC 1.0 ds/m EC 8.0 ds/m I-Core course -Plant Functional
77 Figure 2. Root length (a), root count (b) of tomato under non-saline & saline water irrigation I-Core course -Plant Functional Values are means of sixteen replicates and standard error bars. Asterix mark indicates significant difference between treatments of the same time interval (P =0.05, Student s Phenotyping, t-test) 2018
78 Figure 3. Average root diameter (a) and rooting depth (b) of tomato under non-saline & saline water irrigation Values are means of sixteen replicates and standard error bars. Asterix mark indicates significant difference between treatments of the same time interval (P =0.05, Student s t-test) I-Core course -Plant Functional
79 Figure 4. Changes in root length density (mm/mm 2 ) under non-saline & saline water irrigation Values are means of sixteen replicates and standard error bars. Asterix mark indicates significant difference between treatments of the same depth (P =0.05, Student s t-test) I-Core course -Plant Functional
80 Objectives To develop a new automated minirhizotron (AMR) system for continuous root phenotyping To compare the AMR and manual MR (MMR) systems To continuously monitor the roots for better understanding of root system development & root zone processes I-Core course -Plant Functional
81 2.5 cm 1.3 cm Comparison of the two MR systems Automated MR (AMR) Manual MR (MMR) 5.8 cm 1.8 cm 1 mm = 22 pixels 1 mm = 18 pixels I-Core course -Plant Functional
82 An Automated Minirhizotron (AMR) Systems for precise monitoring of root system response I-Core course -Plant Functional
83 Materials and Methods Grafted tomatoes were grown in sandy soil from Sep 2017 to April 2018 at Ramat Negev Experimental Station Israel A completely AMR system monitored the root system For validation, a regular MMR system (BTC100, Bartz Technology Corporation) was used to collect images Measurements including root length, root count, root distribution were analyzed & compared I-Core course -Plant Functional
84 ARNOLD 1 EC 5 EC A M A M RASISTER 1 EC 5 EC A M A M LURKA 1 EC 5 EC A M A M Comparison of two minirhizotron systems on root system of grafted tomatoes under saline water irrigation A: Automated MR (AMR) M: Manually operated MR (MMR) I-Core course -Plant Functional
85 Figure 1. Simple linear regression of root distribution (%) (length over count basis) by two minirhizotron systems (AMR: Automated MR & MMR: Manually operated MR) in grafted tomatoes I-Core course -Plant Functional
86 R root length by AMR Whether the automated MR camera captures salinity response? R root length by MMR Figure 2. Relative root length (Relative to self grafted) comparison by automated MR (AMR) system and Manual MR (MMR) system on salinity stress response of grafted rootstocks I-Core course -Plant Functional
87 Root length (mm/cm 2 ) by AMR 0-10 cm cm cm cm cm cm Root length (mm/cm 2 ) by MMR Figure 3. Root length comparison of two MR systems (AMR: Automated MR & MMR: Manually operated MR system) on salinity stress response of grafted rootstocks I-Core course -Plant Functional
88 Continuous root monitoring by AMR camera for better understanding of root system development & root zone processes I-Core course -Plant Functional
89 Continuous root monitoring test at net house grown grafted tomatoes MMR AMR MMR Figure 5. Continuous root phenotyping of tomato using an automated MR (AMR) & Manual MR (MMR) system I-Core course -Plant Functional
90 Multiple criteria evaluation of AMR and MMR systems Criteria Automated MR Manual MR Image quality High High Image size and pixels 5.8 cm width 1 mm = 22 pixels 1.8 cm width 1 mm = 18 pixels Time taken for 1 minirhizotron tube 5 min 10 min Cost 1,000 USD 20,000 USD Usage in field Easy/Automated Difficult/Manual/2 persons Accessories Wi-Fi Yes/battery and PC I-Core course -Plant Functional
91 I-Core course -Plant Functional
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93 I-Core course -Plant Functional Gambia '05, Seminar
94 Number of roots per 10 cm as measured with the minirhizotron Wheat study I-Core course -Plant Functional Gambia '05, Seminar
95 I-Core course -Plant Functional Gambia '05, Seminar
96 I-Core course -Plant Functional Gambia '05, Seminar
97 I-Core course -Plant Functional Gambia '05, Seminar
98 I-Core course -Plant Functional
99 צינור מינירייזוטרון שרוול I-Core course -Plant Functional
100 I-Core course -Plant Functional Gambia '05, Seminar
101 Roots and shoots are interdependent. Limitations for one impose limitations on the other. Heavy grazing can decrease the root mass of grass or severe restriction of the root system depresses shoot growth. There are feedback mechanisms for maintaing shoots and roots in relation to one another.
102 During water stress the translocation of photosynthates to the various organs of the plant will change: Water stress generally reduces shoot expansion at a mild level of stress. Roots will receive photosynthates from shoots and elongate new root material wherever soil conditions are favorable.
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105 The timing of water limitation relative to the developmental status of the plant affects allocation patterns: Water stress that occurs during early growth phases causes large shifts in Root/Shoot ratio. In contrast, water stress during reproductive phases (late in the growing season) has little to no effect on the Root/Shoot ratio, but flowering and seed set are reduced or fruit abortion is increased.
106 Eventually, However, when stresses become more severe, root growth can be impacted by the effects of shoot stresses on carbohydrate supply.
107 The next generation: The GFP Minirhizotron In most cases there are roots of several plant in the soil at the same time. The most known cases the combination of weeds and crop but also in other cases there are several root systems growing togeter. I-Core course -Plant Functional
108 One of the more complicates challenges is to distinguish between these different root systems. In some cases its relatively easy - Different root color or root hairs in one crop and no root hairs in the other. In many cases its almost impossible and different methods have to be used. I-Core course -Plant Functional
109 A few years ago a system was developped in the University of Zurich (ETH) by a group of researchers. They developed a modified minirhizotron camera with special filters that enable the user to see the roots of plants with Green Fluorescent Proteins (GFP) at a different color (green) while the roots of the NON GFP will be seen only when the user uses a white regular light sourse. I-Core course -Plant Functional
110 The idea is to take picture with the Minirhizotron system twice: First with white light when you see ALL roots (of the GFP plants and of the NON GFP plants). Second- with the UV light and filters, then you see ONLY the GFP plants. by Subtracting the TOTAL nubmer of roots from the GFP roots, we know what is the number of roots of each of the crops. I-Core course -Plant Functional
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The Hidden Half of the Plant ZACK
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