Response of soybean genotypes to boron,zinc and manganese deficiency in tissue culture

The effect of boron (B), zinc (Zn), and manganese (Mn) deficiency in the medium on eight soybean genotypes was evaluated using friable callus after a 21 day growth period. Of the three micronutrients evaluated, Zn and B deficiencies had the greatest effect on callus weight, while Mn had only a slight effect. Despite this, significant differences in callus weight reduction of the eight genotypes tested were observed on the three different media. These results indicate that genotypic variation for response to B, Zn, and Mn deficiency is present in soybean at the cellular level. If the resistant genotypes identified in this study are also resistant at the whole plant level, then they could be used in soils which are deficient in the above micronutrients.     

In vitro selection of barley and wheat for resistance against Helminthosporium sativum

Summary Calli derived from immature embryos of barley and wheat genotypes were screened for their resistance to purified culture filtrate produced by the fungus Helminthosporium sativum P.K. and B. Two selection methods were used: a continuous method in which four cycles of selection were performed one after another on toxic medium and a discontinuous method in which a pause on non-toxic medium was given after the second or third cycle of selection. The latter was superior as it allowed the calli to regain their regeneration ability. About 3,000 calli from two barley genotypes and 2,000 from two wheat genotypes were used for selection. The selection with the pathotoxins resulted in 6% to 17% surviving calli. Toxin tolerant callus lines of barley were characterised by protein isozymes. Zymograms showed one more isozyme than with the unselected sensitive callus. Barley and wheat plants have been regenerated from callus lines surviving the toxin treatment and in vivo testing against pathogen revealed that the majority of these plants were less sensitive.     

Screening for boron tolerance in wheat (T. aestivum) by solution culture in filter paper

A new screening technique for tolerance to high concentrations of boron, namely a filter paper technique, and a soil experiment were compared to investigate the response of wheat genotypes known to differ in tolerance to high concentrations of boron.Under high boron concentrations in filter papers, the more tolerant genotypes had significantly longer roots than those of the more sensitive genotypes. There was no significant correlation between the root lengths at the control treatment and the other three boron treatments (50, 100, 150 mg B L^-1). Thus, the differences in root lengths at the high boron treatments could not be attributed to inherent differences in root growth but to the genetic variation in response to high boron concentrations among varieties.Root lengths at the three boron treatments in filter papers were highly significantly correlated with the three characters determined for plants grown in soil containing high levels of boron, namely the concentrations of boron in the shoots, plant dry weight and plant symptoms, indicating that root length could be used as a selection criterion in a genetic study or breeding program for boron tolerance.Department of Plant Science, Roseworthy Campus, University of Adelaide     

Possible Involvement of Cytokinin in Nitrate-mediated Root Growth in Maize

Response of root system architecture to nutrient availability in soils is an essential way for plants to adapt to soil environments. Nitrate can affect root development either as a result of changes in the external concentration, or through changes in the internal nutrient status of the plant. Nevertheless, less is known about the physiological mechanisms. In the present study, two maize (Zea mays L.) inbred lines (478 and Wu312) were used to study a possible role of cytokinin in nitrate-mediated root growth in nutrient solutions. Root elongation of 478 was more sensitive to high nitrate supply than that of Wu312. Medium high nitrate (5 mM) inhibited root elongation in 478, while, root elongation in Wu312 was only inhibited at high NO ₃ ⁻ supply (20 mM). Under high nitrate supply, the root elongation zone in 478 became swollen and the site of lateral root elongation was close towards the root tip. Both of the phenomena are typical of root growth induced by exogenous cytokinin treatments. Correspondingly, zeatin and zeatin nucleotide (Z + ZR) concentrations were increased at higher nitrate supply in 478, whereas they were constant in Wu312. Furthermore, exogenous cytokinin 6-benzylaminopurine (6-BA) completely reversed the stimulatory effect of low nitrate on root elongation. Therefore, it is supposed that the inhibitory effect of high concentration of nitrate on root elongation is, at least in part, mediated by increased cytokinin level in roots. High nitrate supply may have negative influences on root apex activity by affecting cytokinin metabolism so that root apical dominance is weakened and, therefore, root elongation is suppressed and lateral roots grow closer to the root apex. Nitrate suppressed lateral root elongation in Wu312 at concentration higher than 5 mM. In 478, however, this phenomenon was not significant even at 20 mM nitrate. Although exogenous 6-BA (20 nM) could suppress lateral root elongation as well, the inhibitory effect of high NO ₃ ⁻ concentration of nitrate on lateral root growth cannot be explained by changes in endogenous cytokinin alone.     

Evaluating the role of root citrate exudation as a mechanism of aluminium resistance in maize genotypes

Organic anion exudation by roots as a mechanism of aluminium (Al) resistance has been intensively studied lately. In the present study, we evaluated qualitative and quantitative aspects of root exudation of organic anions in maize genotypes of distinct sensitivity to Al in response to Al exposure. Maize seedlings were grown axenically in nutrient solution and root exudates were collected along the whole seminal root axis for a short period (4 h) using a divided-root-chamber technique. In root exudates collected from 10-mm long root apices, citrate accounted for 67% of the total organic anions found, followed by malate (29%), trans-aconitate (3%), fumarate (<1%), and cis-aconitate (1%). Rates of citrate exudation from root apices of two genotypes with differential resistance to Al were consistently higher in the Al resistant one, differing by a factor of 1.7 – 3.0 across a range of external Al concentrations. Furthermore, relative Al resistance of eight maize genotypes correlated significantly well with their citrate exudation rate measured at 40 M Al. Higher exudation rates were accompanied by a less inhibited root elongation. The exudation of citrate along the longitudinal axis of fully developed seminal roots showed a particular pattern: citrate was exuded mainly in the regions of root apices, either belonging to the main root or to the lateral roots in the most basal part of the main root. The involvement of citrate in a mechanism of Al resistance is evaluated in terms of protection of the root from the effects of excess Al on root elongation and on nutrient uptake along a root axis showing distinct sites of citrate exudation.     

Auxin-induced inhibition of lateral root initiation contributes to root system shaping in Arabidopsis thaliana

The hormone auxin is known to inhibit root elongation and to promote initiation of lateral roots. Here we report complex effects of auxin on lateral root initiation in roots showing reduced cell elongation after auxin treatment. In Arabidopsis thaliana, the promotion of lateral root initiation by indole-3-acetic acid (IAA) was reduced as the IAA concentration was increased in the nanomolar range, and IAA became inhibitory at 25 nM. Detection of this unexpected inhibitory effect required evaluation of root portions that had newly formed during treatment, separately from root portions that existed prior to treatment. Lateral root initiation was also reduced in the iaaM-OX Arabidopsis line, which has an endogenously increased IAA level. The ethylene signaling mutants ein2-5 and etr1-3, the auxin transport mutants aux1-7 and eir1/pin2, and the auxin perception/response mutant tir1-1 were resistant to the inhibitory effect of IAA on lateral root initiation, consistent with a requirement for intact ethylene signaling, auxin transport and auxin perception/response for this effect. The pericycle cell length was less dramatically reduced than cortical cell length, suggesting that a reduction in the pericycle cell number relative to the cortex could occur with the increase of the IAA level. Expression of the DR5:GUS auxin reporter was also less effectively induced, and the AXR3 auxin repressor protein was less effectively eliminated in such root portions, suggesting that decreased auxin responsiveness may accompany the inhibition. Our study highlights a connection between auxin-regulated inhibition of parent root elongation and a decrease in lateral root initiation. This may be required to regulate the spacing of lateral roots and optimize root architecture to environmental demands.     

Gravitropic response growth angle and vertical distribution of roots of wheat (Triticum aestivum L.)

Recent work on root distribution, growth angles and gravitropic responses in Japanese cultivars of winter wheat are reviewed. Vertical distribution of roots, which influences the environmental stress tolerance of plants, was observed in the 12 Japanese cultivars in the field. The root depth index (RDI: the depth at which 50% of the root length has been reached) differed among the cultivars at the stem elongation stage. Since the RDI was closely related to the growth angle of seminal roots obtained in a pot experiment, it was assumed that growth angle is useful for predicting vertical root distribution among wheat genotypes. Gravitropic responses of the primary seminal root of 133 Japanese wheat cultivars assessed by measuring the growth angle in agar medium, were larger in the northern Japanese cultivars and smaller in the southern ones. It was also found that the geographical variation resulted from the wheat breeding process, i.e. genotypes with limited gravitropic responses of roots had been selected in the southern part of Japan where excessive soil moisture is one of the most serious problems.     

Evaluation of Durum Wheat Genotypes for Resistance against Root Rot Disease Caused by Moroccan Fusarium culmorum Isolates

Fusarium culmorum is one of the most important causal agents of root rot of wheat. In this study, 10 F. culmorum isolates were collected from farms located in five agro-ecological regions of Morocco. These were used to challenge 20 durum wheat genotypes via artificial inoculation of plant roots under controlled conditions. The isolate virulence was determined by three traits (roots browning index, stem browning index, and severity of root rot). An alpha-lattice design with three replicates was used, and the resulting ANOVA revealed a significant (P < 0.01) effect of isolate (I), genotype (G), and G × I interaction. A total of four response types were observed (R, MR, MS, and S) revealing that different genes in both the pathogen and the host were activated in 53% of interactions. Most genotypes were susceptible to eight or more isolates, while the Moroccan cultivar Marouan was reported resistant to three isolates and moderately resistant to three others. Similarly, the Australian breeding line SSD1479-117 was reported resistant to two isolates and moderately resistant to four others. The ICARDA elites Icaverve, Berghisyr, Berghisyr2, Amina, and Icaverve2 were identified as moderately resistant. Principal component analysis based on the genotypes responses defined two major clusters and two sub-clusters for the 10 F. culmorum isolates. Isolate Fc9 collected in Khemis Zemamra was the most virulent while isolate Fc3 collected in Haj-Kaddour was the least virulent. This work provides initial results for the discovery of differential reactions between the durum lines and isolates and the identification of novel sources of resistance.     

Aluminium-induced changes in the surface and micropore properties of wheat roots: a study using the water vapor adsorption-desorption technique

The geometric and energetic characteristics of root surfaces of two wheat (Triticum L.) varieties, Al tolerant (Inia 66/16) and Al sensitive (Henika), were estimated from experimental water vapor adsorption–desorption data. Roots stressed for around 1 week at pH 4 without and with a toxic aluminium level (0.741 mol m^–3) were studied at the tillering and shooting stages. Roots grown continuously at pH 7 were taken as control. The surface properties of the pH 4 stressed roots were apparently the same as those of the control roots whatever the root age. For the roots of both varieties, the surface area and total micropore volume increased markedly after aluminium treatment. The average micropore radius increased significantly for the sensitive wheat, whereas it increased only slightly for the resistant one. Under Al treatment the number of large pores increased while small pores were fewer for both plants, indicating a possible alteration of the build-up of root tissue. The root surface pores were fractal. The fractal dimension of the sensitive wheat roots decreased under Al treatment, whereas for the resistant wheat this remained apparently unchanged. The adsorption energy distribution functions had different shapes for the sensitive and the resistant wheat varieties: the sensitive variety had greater number of high energy adsorption centers, which implies that the root tolerance on Al stress may be connected with lower polarity of the surface.     

Root growth analysis: An underutilised approach to understanding aluminium rhizotoxicity

Despite numerous published studies, the mechanistic bases for Al rhizotoxicity and for intraspecific differences in Al tolerance remain elusive. Classical methods of growth analysis offer a means to describe quantitatively the onset of root growth inhibition in response to an Al challenge, and any subsequent recovery in growth rates (i.e. acclimation). Such information could help elucidate tolerance mechanisms (i.e. constitutive vs. inducible). Despite this potential utility, however, published data of high accuracy and precision are few, especially with wheat (Triticum aestivum L.). In this paper, a conceptual framework for analysing root growth responses to Al is presented. Specially designed tanks and a computerised video analysis system were used to monitor root elongation in a simple medium (1 mM CaCl₂, pH 4.3) containing varied Al concentrations. Root lengths of five wheat cultivars were measured every 2 to 5 h for 48 h after Al challenge. Growth rates typically exhibited a brief (1–2 h) lag phase prior to a sharp decline, followed by a recovery phase of 12 h duration. At low levels of Al (i.e. sufficient to cause a 10% reduction in net elongation) post-recovery growth rates generally equalled those of control roots, but were significantly reduced at Al concentrations yielding a 40 to 50% reduction in net elongation. All cultivars exhibited similar inhibition-acclimation response patterns and these responses did not explain differences in sensitivity to Al. Nonetheless, solution Al concentrations required to elicit the same growth responses varied by 20-fold across the five cultivars, consistent with the notion that root exclusion of Al is a significant component of differential Al tolerance in wheat.     

Genetic Analysis of Maize Root Characteristics in Response to Low Nitrogen Stress

Under low-input cropping systems, nitrogen (N) can be a limiting factor in plant growth and yield. Identifying genotypes that are more efficient at capturing limited N resources and the traits and mechanisms responsible for this ability is important. Root trait has a substantial influence on N acquisition from soils. Nevertheless, inconsistencies still exist as to the effect of low N on root length and its architecture in terms of lateral and axial roots. For maize, a crop utilizing heterosis, little is known about the relationship between parents and their crosses in the response of root architecture to N availability. Here 7 inbred maize lines and 21 of their crosses created by diallel mating were used to study the effect of N stress on root morphology as well as the relationship between the inbreds and their crosses. With large genotypic differences, low N generally suppresses shoot growth and increases the root to shoot ratio with or without increasing root biomass in maize. Maize plants responded to N deficiency by increasing total root length and altering root architecture by increasing the elongation of individual axial roots and enhancing lateral root growth, but with a reduction in the number of axial roots. Here, the inbreds showed weaker responses in root biomass and other root parameters than their crosses. Heterosis of root traits was significant at both N levels and was attributed to both the general combining ability (GCA) and special combining ability (SCA). Low N had substantial affects on the pattern of heterosis, GCA and SCA affects on root traits for each of the crosses suggesting that selection under N stress is necessary in generating low N-tolerant maize genotypes.     

The Effects of Aluminium on the Seed Germination and Seedling Growth in Hibiscus moscheutos (Malvaceae) and Wheat (Triticum aestivaum,Gramineae)

The effects of AlCl₃ on Hibisucs moscheutos seed germination and growth were investigated to evaluate its hardiness to Aluminum (Al), in compassion with Carazinho, a wheat genotype tolerant to Al, and Egret, agenotype sensitiveto Al . For H. moscheutos and two wheat genotypes, our results indicated that germination was insensitive to AlCl₃ until at 500μmol/L. AlCl₃ of 50μmol/L inhibited the elongation of primary and lateral roots significantly, but had less effect on the number of lateral roots. There was no difference of root elongation between the H. moscheutos and wheat, but the lateral roots of H. moscheutos were more tolerant to Al than that of wheat genotypes. Under AlCl₃ of 50μmol/L, the reduction of root biomass was significant in wheat genotypes, but not in H. moscheutos, comparing to their control, suggesting that H. moscheutos is more tolerant to Al than the two wheat genotypes, and that Al has different effects on the growth of primary and lateral roots in both H. moscheutos and wheat .     

Short root mutant of Lotus japonicus with a dramatically altered symbiotic phenotype

Legume plants carefully control the extent of nodulation in response to rhizobial infection. To examine the mechanism underlying this process we conducted a detailed analysis of the Lotus japonicus hypernodulating mutants, har1-1, 2 and 3 that define a new locus, HYPERNODULATION ABERRANT ROOT FORMATION (Har1), involved in root and symbiotic development. Mutations in the Har1 locus alter root architecture by inhibiting root elongation, diminishing root diameter and stimulating lateral root initiation. At the cellular level these developmental alterations are associated with changes in the position and duration of root cell growth and result in a premature differentiation of har1-1 mutant root. No significant differences between har1-1 mutant and wild-type plants were detected with respect to root growth responses to 1-aminocyclopropane1-carboxylic acid, the immediate precursor of ethylene, and auxin; however, cytokinin in the presence of AVG (aminoetoxyvinylglycine) was found to stimulate root elongation of the har1-1 mutant but not the wild-type. After inoculation with Mesorhizobium loti, the har1 mutant lines display an unusual hypernodulation (HNR) response, characterized by unrestricted nodulation (hypernodulation), and a concomitant drastic inhibition of root and shoot growth. These observations implicate a role for the Har1 locus in both symbiotic and non-symbiotic development of L. japonicus, and suggest that regulatory processes controlling nodule organogenesis and nodule number are integrated in an overall mechanism governing root growth and development.     

Genotype and fungicide effects on late-season root growth of winter wheat

The aim of this work was to investigate differences among genotypes in post-anthesis root growth and distribution of modern UK winter wheat cultivars, and the effects of fungicide applications. Post-anthesis root growth of up to six cultivars of winter wheat (Triticum aestivum L.), given either one or three applications of fungicide, was studied in field experiments during two seasons. Total root mass remained unchanged between GS63 (anthesis) and GS85, but root length increased significantly from 14.7 to 31.4 km m⁻² in one season. Overall, there was no evidence for a decline in either root mass or length during grain filling. Root mass as a proportion of total plant mass was about 0.05 at GS85. There were significant differences among cultivars in root length and mass especially below 30 cm. Malacca had the smallest root length and Savannah the largest, and Shamrock had a significantly larger root system below 40 cm in both seasons. Fungicide applied at ear emergence had no significant effect on root mass in either season but increased root length (P<0.01) in the more disease-prone season. By maintaining a green canopy for longer, fungicide applied at flag leaf emergence may have resulted in delayed senescence of the root system and contributed to the post-anthesis maintenance of root mass and length. Section Editor: R. W. Bell     

Effects of Leaf Wetness Duration and Inoculum Level on Resistance of Wheat Genotypes to Pyrenophora tritici-repentis

Percent leaf necrosis and lesion length on wheat genotypes increased markedly with increasing duration of leaf wetness (up to 24h or 48 h) following inoculation with Pyrenophora tritici-repentis. A long wetting duration favoured less disease development on resistant (Fink's'), and moderately resistant (Bon/YR/3/F3570//KAL/BB) genotypes than on susceptible Glenlea. No significant difference in per cent necrosis was detected among the upper three leaf positions within a genotype. A long wetness duration had a varying effect on the resistance of wheat genotypes, depending upon the inoculum level. Increasing the inoculum level along with the leaf wetness period increased the per cent leaf necrosis on all three wheat genotypes tested. However, the ranking of the genotype for resistance did not alter even after prolonged duration of leaf wetness (up to 96 h) and/or high inoculum level (12000 conidia/ml water). Various post-inoculation wet-periods in combination with high conidia concentrations in inoculum should be used in identifying highly resistant germplasm in breeding populations at the seedling stage of the wheats.     

Effects of Leaf Wetness Duration and Inoculum Level on Resistance of Wheat Genotypes to Pyrenophora tritici-repentis

Percent leaf necrosis and lesion length on wheat genotypes increased markedly with increasing duration of leaf wetness (up to 24h or 48 h) following inoculation with Pyrenophora tritici-repentis. A long wetting duration favoured less disease development on resistant (Fink's'), and moderately resistant (Bon/YR/3/F3570/KAL/BB) genotypes than on susceptible Glenlea. No significant difference in percent necrosis was detected among the upper three leaf positions within a genotype. A long wetnessduration had a varying effect on the resistance of wheat genotypes, depending upon the inoculum level. Increasing the inoculum level along with the leaf wetness period increased the per cent leaf necrosis on all three wheat genotypes tested. However, the, ranking of the genotype for resistance did not alter even after prolonged duration of leaf wetness (up to 96 h) and/or high inoculum level (12000 conidia/ml water). Various post-inoculation wet-periods in combination with high conidia concentrations in inoculum should be used in identifying highly resistant germplasm in breeding populations at the seedling stage of the wheats.     

Effects of applying stem-shortening plant growth regulators to leaves on root elongation by seedlings of wheat, oat and barley: mediation by ethylene

Several plant growth regulators (PGRs) commonly used in practicalfarming to restrict shoot height and control lodging were examined for theirimpact on root growth in naturally short or tall cultivars of barley (cvs.Kymppi and Saana), oat (cvs. Veli and Pal), and wheat (cvs. Mahti and Tjalve).The possible involvement of ethylene in the responses was also examined. Shootswere sprayed at the two-leaf stage with the gibberellin biosynthesis inhibitorsCycocel (chlormequat chloride) (CCC) or Moddus (Trinexapac-ethyl) (TE), or withthe ethylene-releasing agent Cerone (ethephon) (ETH) at 0, 0.1, 1, 10 or 50times the recommended agricultural rate (RR). Root elongation and ethyleneproduction by roots or shoots were unaffected by CCC at all application ratesorby TE at ×0.1 or ×1.0 RR. At ×10 and ×50 RR, TE wasinhibitory to root extension but did not increase ethylene biosynthesis bytheseroots or the shoots. ETH at ×0.1 or ×1.0 RR did not affect rootextension or ethylene production in roots or shoots. At all higher rates ofapplication ETH stimulated ethylene production strongly in shoots and roots ofall three species, while root elongation was retarded severely in barley,moderately in oat and only slightly in wheat. These differences in elongationresponse are attributed to differences in sensitivity to ethylene released byethephon. Accordingly, root elongation in wheat was only slightly affected whenethylene gas was supplied at concentrations up to 100 ppm for 3d. In contrast, root elongation in barley was strongly inhibitedbyethylene, with oat demonstrating an intermediate responsiveness.     

Heterogeneous nutrient supply promotes maize growth and phosphorus acquisition: additive and compensatory effects of lateral roots and root hairs

Background and AimsRoot proliferation is a response to a heterogeneous nutrient distribution. However, the growth of root hairs in response to heterogeneous nutrients and the relationship between root hairs and lateral roots remain unclear. This study aims to understand the effects of heterogeneous nutrients on root hair growth and the trade-off between root hairs and lateral roots in phosphorus (P) acquisition.MethodsNear-isogenic maize lines, the B73 wild type (WT) and the rth3 root hairless mutant, were grown in rhizoboxes with uniform or localized supply of 40 (low) or 140 (high) mg P kg⁻¹ soil.ResultsBoth WT and rth3 had nearly two-fold greater shoot biomass and P content under local than uniform treatment at low P. Significant root proliferation was observed in both WT and rth3 in the nutrient patch, with the WT accompanied by an obvious increase (from 0.7 to 1.2 mm) in root hair length. The root response ratio of rth3 was greater than that of WT at low P, but could not completely compensate for the loss of root hairs. This suggests that plants enhanced P acquisition through complementarity between lateral roots and root hairs, and thus regulated nutrient foraging and shoot growth. The disappearance of WT and rth3 root response differences at high P indicated that the P application reduced the dependence of the plants on specific root traits to obtain nutrients.ConclusionsIn addition to root proliferation, the root response to a nutrient-rich patch was also accompanied by root hair elongation. The genotypes without root hairs increased their investment in lateral roots in a nutrient-rich patch to compensate for the absence of root hairs, suggesting that plants enhanced nutrient acquisition by regulating the trade-off of complementary root traits.     

Exogenous auxin effects on growth and phenotype of normal and hairy roots of Pueraria lobata (Willd.) Ohwi

Pueraria lobata hairy roots have faster elongationand more branches than normal roots. The responses of hairy roots and normalroots to treatment with three auxins, indole-3-acetic acid (IAA),indole-3-butyric acid (IBA), and naphthalene acetic acid (NAA) were different.In normal roots, all three auxins strongly stimulated lateral root formation atall tested concentrations. Responses to IAA and IBA in primary root growth andlateral root elongation were similar and depended on concentration; promotionat0.1 M, no effect at 1.0 M, and inhibition at2.5 M. In hairy roots, lateral root formation varied inresponseto the different auxins, i.e. depressed by NAA, unaffected by IAA, and promotedby IBA. Primary root growth was slightly inhibited by IBA and was unaffected byIAA. However, mean lateral root length was reduced in response to IAA and IBA.Only NAA exerted strong inhibition on primary and lateral root elongation inboth root types. The similar free IAA and conjugated IAA content but quitedifferent basal ethylene production and biosynthesis in hairy and normal rootssuggested different mechanisms of response to exogenous auxins in the two roottypes.     

Evaluating the Madeiran wheat germplasm for aluminum resistance using aluminium-induced callose formation in root apices as a marker

Aluminum (Al) resistance of 57 Madeiran wheat cultivars was evaluated using callose content in root tips and root elongation as markers. Al induced callose formation was a very sensitive indicator of Al damage detecting wide range of genotypic differences existing in the Madeiran wheat germplasm. A weak, yet positive correlation (R²=0.285, P<0.05) between callose content and root elongation was found.