Taiwen Yong

Transcriptome Analysis of Shade-Induced Inhibition on Leaf Size in Relay Intercropped Soybean

Multi-species intercropping is a sustainable agricultural practice worldwide used to utilize resources more efficiently. In intercropping systems, short crops often grow under vegetative shade of tall crops. Soybean, one important legume, is often planted in intercropping. However, little is known about the mechanisms of shade inhibition effect on leaf size in soybean leaves at the transcriptome level. We analyzed the transcriptome of shaded soybean leaves via RNA-Seq technology. We found that transcription 1085 genes in mature leaves and 1847 genes in young leaves were significantly affected by shade. Gene ontology analyses showed that expression of genes enriched in polysaccharide metabolism was down-regulated, but genes enriched in auxin stimulus were up-regulated in mature leaves; and genes enriched in cell cycling, DNA-replication were down-regulated in young leaves. These results suggest that the inhibition of higher auxin content and shortage of sugar supply on cell division and cell expansion contribute to smaller and thinner leaf morphology, which highlights potential research targets such as auxin and sugar regulation on leaves for crop adaptation to shade in intercropping.

Growth and photosynthetic responses of soybean seedlings to maize shading in relay intercropping system in Southwest China

Intercropping, a traditional and worldwide cropping practice, has been considered as a paradigm of sustainable agriculture based on complementary mechanisms among different crop species. Soybean (Glycine max) is widely relay intercropped about 60 days before maize (Zea mays) harvest in Southwest China. However, shade caused by tall maize plants may be a limiting factor for soybean growth at a seedling stage. In field research, we studied the ecophysiological responses of two widely cultivated soybean varieties [Gongqiudou494-1 (GQD) and Gongxuan 1 (GX)] to maize shading in a relay intercropping system (RI) compared with monocropped soybean plants (M). Our results showed that soybean seedlings intercropped with maize exhibited significantly downregulated net photosynthetic rate (PN) (−38.3%), transpiration rate (−42.7%), and stomatal conductance (−55.4%) due to low available light. The insignificant changes in intercellular CO2 concentration and the maximum efficiency of PSII photochemistry suggested that the maize shading-induced depressions in PN were probably caused by the deficiency of energy for carbon assimilation. The significantly increased total chlorophyll (Chl) content (+27.4%) and Chl b content (+52.2%), with lowered Chl a/b ratios (−20.5%) indicated soybean plants adjusted their light-harvesting efficiency under maize shading condition. Biomass and leaf area index (LAI) of seedlings under RI decreased significantly (−78.7 and −71%, respectively) in comparison with M. Correlation analysis indicated the relative reduction in biomass accumulation was caused by the decline in LAI rather than PN, it affected negatively the final yields of soybean (32.8%). Cultivar-specific responses to maize shading were observed in respects of LAI, biomass, and grain yield. It indicated that GX might be a better cultivar for relay intercropping with maize in Southwest China.

Karrikins delay soybean seed germination by mediating abscisic acid and gibberellin biogenesis under shaded conditions

Karrikins (KAR) are a class of signal compounds, discovered in wildfire smoke, which affect seed germination. Currently, numerous studies have focused on the model plant Arabidopsis in the KAR research field, rather than on crops. Thus the regulatory mechanisms underlying KAR regulation of crop seed germination are largely unknown. Here, we report that KAR delayed soybean seed germination through enhancing abscisic acid (ABA) biosynthesis, while impairing gibberellin (GA) biogenesis. Interestingly, KAR only retarded soybean seed germination under shaded conditions, rather than under dark and white light conditions, which differs from in Arabidopsis. Phytohormone quantification showed that KAR enhanced ABA biogenesis while impairing GA biosynthesis during the seed imbibition process, and subsequently, the ratio of active GA4 to ABA was significantly reduced. Further qRT-PCR analysis showed that the transcription pattern of genes involved in ABA and GA metabolic pathways are consistent with the hormonal measurements. Finally, fluridone, an ABA biogenesis inhibitor, remarkably rescued the delayed-germination phenotype of KAR-treatment; and paclobutrazol, a GA biosynthesis inhibitor, inhibited soybean seed germination. Taken together, these evidences suggest that KAR inhibit soybean seed germination by mediating the ratio between GA and ABA biogenesis.

Fungal Diversity in Field Mold-Damaged Soybean Fruits and Pathogenicity Identification Based on High-Throughput rDNA Sequencing

Continuous rain and an abnormally wet climate during harvest can easily lead to soybean plants being damaged by field mold (FM), which can reduce seed yield and quality. However, to date, the underlying pathogen and its resistance mechanism have remained unclear. The objective of the present study was to investigate the fungal diversity of various soybean varieties and to identify and confirm the FM pathogenic fungi. A total of 62,382 fungal ITS1 sequences clustered into 164 operational taxonomic units (OTUs) with 97% sequence similarity; 69 taxa were recovered from the samples by internal transcribed spacer (ITS) region sequencing. The fungal community compositions differed among the tested soybeans, with 42 OTUs being amplified from all varieties. The quadratic relationships between fungal diversity and organ-specific mildew indexes were analyzed, confirming that mildew on soybean pods can mitigate FM damage to the seeds. In addition, four potentially pathogenic fungi were isolated from FM-damaged soybean fruits; morphological and molecular identification confirmed these fungi as Aspergillus flavus, A. niger, Fusarium moniliforme, and Penicillium chrysogenum. Further re-inoculation experiments demonstrated that F. moniliforme is dominant among these FM pathogenic fungi. These results lay the foundation for future studies on mitigating or preventing FM damage to soybean.

Characteristics of Nitrogen Uptake, Use and Transfer in a Wheat-Maize-Soybean Relay Intercropping System

Abstract Intercropping and relay intercropping systems, which significantly improve land use efficiency, are used worldwide to increase crops yield. The wheat-maize-soybean relay intercropping system has been widely employed by famers in Southwestern China for years, but the detailed mechanisms through which the nitrogen fertilizer use efficiency reach the high level in this system remain unclear. In the present study, two separate pot experiments were performed by 15 N isotope dilution (ID) labeling and direct 15N foliar feeding (FF) assays, and a solid barrier was employed to prevent the roots intergrowth and N movement among crops in the first experiment, using no barrier as the control. The results showed that, under the no-barrier condition, the grain yields, 15N uptake and 15N recovery efficiency of wheat and maize were significantly increased, but those measures in soybean were decreased compared to the solid barrier condition. Furthermore, bi-directional N transfer was detected during the co-growing stage of crops, the amount (Ntransfer) and percentage (%NT) of 15N transferred varied significantly with the fertilizer-N rate, and the maximum reached at 150 – 300 kg N ha–1 level. The Ntransfer from maize to wheat was 16.1% – 163.0% higher than that from wheat to maize; the Ntransfer from soybean to maize was 1.7 – 6.0 times higher than those from maize to soybean, while the %NT from soybean to maize were 6.7 – 22.2 times higher than those from maize to soybean. Conclusively, this study revealed that the interaction of the roots among crops significantly increased the uptake efficiency and recovery efficiency, and further, the positive N competition and bi-directional N transfer of each crops were the main contributors to improve the N use efficiency in the wheat-maize-soybean relay intercropping system.

Salt Stress Represses Soybean Seed Germination by Negatively Regulating GA Biosynthesis While Positively Mediating ABA Biosynthesis

Soybean is an important and staple oilseed crop worldwide. Salinity stress has adverse effects on soybean development periods, especially on seed germination and post-germinative growth. Improving seed germination and emergence will have positive effects under salt stress conditions on agricultural production. Here we report that NaCl delays soybean seed germination by negatively regulating gibberellin (GA) while positively mediating abscisic acid (ABA) biogenesis, which leads to a decrease in the GA/ABA ratio. This study suggests that fluridone (FLUN), an ABA biogenesis inhibitor, might be a potential plant growth regulator that can promote soybean seed germination under saline stress. Different soybean cultivars, which possessed distinct genetic backgrounds, showed a similar repressed phenotype during seed germination under exogenous NaCl application. Biochemical analysis revealed that NaCl treatment led to high MDA (malondialdehyde) level during germination and the post-germinative growth stages. Furthermore, catalase, superoxide dismutase, and peroxidase activities also changed after NaCl treatment. Subsequent quantitative Real-Time Polymerase Chain Reaction analysis showed that the transcription levels of ABA and GA biogenesis and signaling genes were altered after NaCl treatment. In line with this, phytohormone measurement also revealed that NaCl considerably down-regulated active GA1, GA3, and GA4 levels, whereas the ABA content was up-regulated; and therefore ratios, such as GA1/ABA, GA3/ABA, and GA4/ABA, are decreased. Consistent with the hormonal quantification, FLUN partially rescued the delayed-germination phenotype caused by NaCl-treatment. Altogether, these results demonstrate that NaCl stress inhibits soybean seed germination by decreasing the GA/ABA ratio, and that FLUN might be a potential plant growth regulator that could promote soybean seed germination under salinity stress.

Responses to shade and subsequent recovery of soya bean in maize-soya bean relay strip intercropping

Abstract In relay intercropping systems, late-planted crops often grow under the shade of the canopy of early-planted tall crops and then transfer to full sunlight after the harvest of the early-planted crops. In order to know the effects of recovery growth of the late-planted soya bean in maize–soya bean relay intercropping, a field experiment was carried out to observe architectural, morphological, physiological and anatomical traits of soya bean plants related to shade and subsequent removal in intercropping before and after maize harvest, respectively. During shade period, soya bean biomass was severely reduced, and stem elongation was stimulated. Typical features of shade grown leaves were found, such as lower LMA (leaf mass per unit area), thinner thickness, higher chlorophyll content, lower chlorophyll a:b ratio. Whole-plant leaf area analysis found that soya bean increased leaf area ratio by adjusting leaf morphology rather than by dry mass allocation. After maize harvest, leaf area and leaf mass increased rapidly, contributing to compensation growth in intercropped soya bean. Meanwhile, physiological and anatomical traits of leaf went back to similar levels as grown in sole cropping. However, stem morphological traits were irreversible after removal of shade. Finally, no difference on seed weight per plant of soya bean was observed between relay intercropping and sole cropping. Based on these findings, we speculated the recovery growth might be the direct determining factor on pod formation in soya bean, and improvement on the capacity of recovery growth could increase yield of relay intercropped soya bean.

PAR Interception and Utilization in Different Maize and Soybean Intercropping Patterns

The crop intercepted photosynthetically active radiation (PAR) and radiation use efficiency (RUE) vary markedly in different intercropping systems. The HHLA (horizontally homogeneous leaf area) and ERCRT (extended row crop radiation transmission) models have been established to calculate the intercepted PAR for intercrops. However, there is still a lack of study on the intercepted PAR and RUE under different intercropping configurations using different models. To evaluate the intercepted PAR and RUE in maize and soybean under different intercropping systems, we tested different strip intercropping configurations (SI1, SI2, and SI3 based on ERCRT model) and a row intercropping configurations (RI based on HHLA model) in comparison to monoculture. Our results showed that the intercepted PAR and RUE of intercropping systems were all higher than those of monoculture. The soybean intercepted PAR in strip intercropping was 1.35 times greater than that in row intercropping. In row intercropping (RI), the lack of soybean intercepted PAR resulted in a significant reduction of soybean dry matter. Therefore, it is not the recommended configuration for soybean. In strip intercropping patterns, with the distance between maize strip increased by 0.2 m, the intercepted PAR of soybean increased by 20%. The SI2 (maize row spacing at 0.4 m and the distance between maize strip at 1.6 m) was the recommended configuration to achieve the highest value of intercepted PAR and RUE among tested strip intercropping configurations. The method of dry matter estimation using intercepted PAR and RUE is useful in simulated experiments. The simulated value was verified in comparison with experimental data, which confirmed the credibility of the simulation model. Moreover, it also provides help in the development of functional-structural plant model (FSPM).

Metabolomic tool to identify soybean [ Glycine max (L.) Merrill] germplasms with a high level of shade tolerance at the seedling stage

The isoflavone profiles of seeds of various soybean genotypes with different levels of shade tolerance at the seedling stage were investigated. High-performance liquid chromatography (HPLC) was used to quantify 12 isoflavones, and the data were analyzed using a multivariate statistical analysis. Combined with field experimental data and an orthogonal partial least-squares discriminant analysis (OPLS-DA), several aglycones (genistein (GE), daidzein (DE), and glycitein (GLE)) were selected and identified as key compounds involved in the shade tolerance of soybean seedlings. Additional correlation analysis and laboratory shading stress experiments with soybean seedlings also confirmed the function of these selected isoflavones, especially GE, in the discrimination of soybean seedlings with different levels of shade tolerance. Furthermore, the structure–antioxidant activity relationships between a range of isoflavones and the plant shade-tolerance mechanism are discussed. Targeted metabolomic analyses of isoflavones could reveal the diversity of shade tolerance in soybean seedlings, thus contributing to the breeding of excellent soybean varieties.

Effect of narrow-row planting patterns on crop competitive and economic advantage in maize-soybean relay strip intercropping system

Abstract Narrow-row planting patterns directly affect crop yield and competition in intercropping systems. A two-year (2012 and 2013) field experiment was conducted to determine the interactive behavior between intercrops in a maize–soybean relay strip intercropping system. Maize plants were planted in different narrow-wide row planting patterns, whereas soybean was planted in wide rows. The total biomass and grain yield of maize increased with increasing maize narrow-row spacing, but the opposite trend was observed for soybean. The aggressivity, competitive ratio, and partial relative crowding advantage values for maize were greater than those for soybean. Moreover, the competitive interaction of the intercrops was affected by the distance between maize and soybean rows. The highest intercrop land equivalent ratio (LER) 1.61 and 1.59 was found in the 40:160 planting pattern (i.e. 40 cm narrow-row spacing and 160 cm wide-row spacing of maize) during 2012 and 2013, respectively. Combined with actual yield loss and LER, the intense intra-specific competition of maize plants reduced the depression for the associated soybeans when the maize narrow-row spacing was less than 30 cm. When the narrow-row spacing was wider than 50 cm, soybean growth was seriously depressed by maize because of the stronger inter-specific competition between maize and soybean. The maximum yield and economic advantage appeared in the 40:160 narrow-wide row planting pattern. Therefore, intercropping advantage may be achieved by changing the row spacing and distance between intercrop rows to coordinate the inter-specific competition between maize and soybean.