Evaluation of root lodging resistance during whole growth stage at the plant level in maize
Experimental design and crop managementField experiments were conducted at Chengyang Agricultural Experimental Station, Qingdao, China (36°18′ 11″/N, 120°21′ 13″/E) in 2019 and 2020. The soil type in the field was brown loam that contained 22.76 g kg−1 organic matter, 82.39 mg kg−1 alkali-hydrolysable N, 25.10 mg kg−1 Olsen-P and 94.89 mg kg−1 exchangeable K. The test cultivars of maize were Jinhai5 with strong lodging resistance and Xundan20 with weak lodging resistance, which were repeated four times in plots laying out in randomized block designs. Plant density was 7.5 plants / m2 with the row spacing of 60 cm. the plot consisted of 8 rows length of 15 m. Two–three seeds per hole were manually sowed at 5 cm on 20 April 2019 and 24 April 2020, and the seedlings were thinned to the target planting density at V2, and harvested on 10 September and 14 September, respectively. Fertilization and irrigation management followed local production practices in maize.Sampling and measurementPlant samples were taken at V8, V12, R1, R2 and R6. Ten typical plants of each tested cultivars were selected to be subjected to mechanical and above-ground morphological measurements at each sampling. The other three maize plants were used to measure morphological traits of roots. Xundan20 was seriously damaged due to the storm in the late stage of maize growth in 2020, resulting in the missing data for physiological maturity.Determination of leaf area vertical distributionLeaf area of expanded leaves each was computed by the coefficient method: Single leaf area = length * width * 0.75. Leaf area for unexpanded leaves was estimated by the leaf weight method. Leaf area per plant was the sum of all individual green leaf areas. Leaf height is the height from the ground to the leaf collar position of maize.Determination of max root side-pulling resistanceSample plants were surrounded with water-proof steel devices inserted into underground, and watered to soil moisture over saturation at one day before mechanical testing. When measured, due to the limited space, all leaves of sample plants are removed in order to improve the measurement accuracy. The defoliated stalks were immobilized by a pair of lengthwise steel clamps to prevent stalks from bending (Fig. 7). After the digital pole dynamometer18 with a 1.5 m long slider and a main unit was linked to the stalks at a height of 80 cm away from the ground, the operator by hand pulled at a slow and uniform speed until the roots were pulled out. Records of load force, declination angle and sensor position were automatically stored in main unit during this operation. The peak value of forces, extracted from records, was taken as the max root side-pulling resistance.Figure 7Schematic diagram for measuring max root side-pulling resistance.Full size imageRoot anti-lodging indexBased on the method of Cui et al.6, the force value comparison is changed to the moment value comparison to calculate root anti-lodging index:$${text{AL}}_{root} = M_{root} / , M_{wind} = F_{root} / , F_{wind}$$
(1)
where M root is the root failure moment, M wind is the wind resultant moment. Root anti-lodging index indicates the ability of plants to resist root lodging. The larger its value is, the stronger the resistance is, and vice versa.$${text{M}}_{root} = F , *d$$
(2)
where F is the max root side-pulling resistance, d is moment arm, i.e., the length of force arm. As a component of root anti-lodging index, the root failure moment represents the ability of the root system to resist lateral pulling. The greater its value is, the better the resistance is, and vice versa.With the base of the stem as the fulcrum,$${text{M}}_{wind} = sum 0.{5}CA_{i} rho V^{2} h_{i}$$
(3)
where C is coefficient of air resistance, ρ is air mass density ,V is the wind speed , Ai is the area of a single leaf , hi is the height of leaf, ∑ represents to sum up over all leaves. C value is set to be 0.219. When encountering wind speed at grade 6 or higher, maize is more prone to lodging. Unless stated explicitly, the following analysis was limited to the upper wind speed for grade 6 wind20.Root morphological traitsThe number and length of all primary nodal roots were measured. Root-soil balls each of two or three tested plants were obtained after lateral root-pulling testing. The images of the three frontal sides, 120 degrees apart from each other, of the root-soil balls were taken using a digital camera. Ball volumes were then evaluated by considering them to be rotationally symmetric. Average volumes were used for further analysis.Single root tensile resistanceRoots after counting the number of nodal roots were used to measure the single root tensile resistance. First, clean the dust off roots. Then, diameters of roots were determined with a vernier caliper. Single root tensile resistance was measured by HF-500 digital push–pull apparatus. Fixed the upper and lower ends of the root, then one end moved slowly and uniformly, the other end was still until the root breaks. The peak tension force displayed by the instrument was taken as the single root tensile resistance.Statistical analysisBased on variance analysis, the Tukey method was used to compare the differences among means. The logarithmic transformation of variables was carried out to improve the homogeneity of error variance if appropriate.The substantive effect or influence of various factors on the response variable can be expressed by effect size of factors, which can be calculated under the framework of variance analysis. Effect size is the proportion of the effect of a certain factor in the total effect, which is a dimensionless number21,22,23.The formula for calculating effect size of factors is:$$omega^{2} = frac{{df_{effect} times left( {MS_{effect} – MS_{error} } right)}}{{SS_{total} + MS_{error} }}$$
(4)
where df is the degree of freedom, MS represents mean square.Two conceptual models were used when dealing with effect size. One model was of components, i.e., taking the logarithm of both sides of Eq. (1):$${text{LOG}}left( {{text{AL}}_{{{text{root}}}} } right) , = {text{ LOG}}left( {{text{M}}_{{{text{root}}}} } right) , + {text{ LOG}}left( {{text{M}}_{{{text{wind}}}} } right)$$
(5)
where LOG denotes logarithmic transformation.The other was the factorial model, i.e.,$${text{factors affecting AL}}_{{{text{root}}}} = {text{ wind grade }} + {text{ cultivar }} + {text{ growth stage}}$$
(6)
Experimental research and field studies on plants including the collection of plant materialThe authors declare that the cultivation of plants and carrying out study in Chengyang Agricultural Experimental Station complies with all relevant institutional, national and international guidelines and treaties.Statement of permissions and/or licenses for collection of plant or seed specimensThe authors declare that the seed specimens used in this study are publicly accessible seed materials and we were given explicit written permission to use them for this research. More