Anti-Rhizoctonia solani activity by Desmos chinensis extracts and its mechanism of action

The objectives of this study were to investigate the mechanisms of action and to evaluate the potential application of Desmos chinensis extracts for controlling rice sheath blight. The dichloromethane extract from D. chinensis demonstrated high antifungal activity against Rhizoctonia solani. The extract was shown to have anti-R. solani activity with minimum inhibitory concentration (MIC) and minimum fungicidal concentration values ranging from 31.2 to 62.5 μg mL⁻¹ and from 62.5 to 500 μg mL⁻¹, respectively. Bioautography on thin-layer chromatography plates demonstrated antifungal activity of the extract with an R_f value of 0.33. A total of 7 compounds, 2 benzoate esters (benzyl benzoate and benzyl 2-hydroxybenzoate), 2 sesquiterpenoids (α-eudesmol and β-eudesmol), 1 aromatic alcohol (benzyl alcohol), 1 aromatic ketone (acetophenone) and 1 diterpenoid (phytol), were detected and identified using gas chromatography–mass spectrometry analysis. Electron micrographs confirmed the effects of the extract on morphological and ultrastructural alterations in the treated fungal cells. The micrographs of the mycelia treated with the extract at 4MIC illustrated aberrant morphology such as shrinkage, partial distortion and globular structures of different sizes along the surface of the mycelia. Damaging membranous structures including disruption of the cell membrane, partial loss of nuclear membranes, and depletion of hyphal cytoplasm and membranous organelles were observed. Foliar application of D. chinensis extract at a concentration of 2 mg mL⁻¹ on rice markedly decreased sheath blight severity. It was concluded that the application of D. chinensis extract can be used as a botanical fungicide to control rice sheath blight.     

Screening of Brown Planthopper Resistant miRNAs in Rice and Their Roles in Regulation of Brown Planthopper Fecundity

MicroRNAs (miRNAs) can participate in plant-insect interactions, which regulate plant defense networks. In this study, we analyzed the miRNA expression profiles of six rice varieties before and after brown planthopper (BPH)-feeding. We identified 45 differentially expressed miRNAs between BPH- susceptible and BPH-resistant rice varieties and 144 miRNAs that responded to BPH-feeding. Thus, miRNAs may be involved in multiple pathways regulating rice defense response against BPH. In addition, we found that the genetic history of rice varieties determined the regulation mode of the miRNA and affected the amounts, types, changing trends and response periods of miRNAs in response to BPH- feeding. To conclude, we scanned seven potential cross-kingdom miRNAs, of which miR5795 may target the vitellogenin gene in BPH, causing a 16.07% reduction in BPH oviposition. The results provide new miRNA information of rice-BPH interactions and BPH-resistant rice variety breeding.     

Anti-fungal action of bioconverted eicosapentaenoic acid (bEPA) against plant pathogens

Bioconverted eicosapentaenoic acid (bEPA), obtained from Pseudomonas aeruginosa PR3, was assessed for its in vitro and in vivo anti-fungal potential. Mycelial growth inhibition of the tested plant pathogens (Rhizoctonia solani, Botrytis cinerea, Fusarium oxysporum, Fusarium solani, Phyptophthora capsici, Sclerotinia sclerotiorum and Colletotrichum capsici) was measured in vitro. bEPA at the concentration of 5 μl/ml inhibited 52–60% fungal mycelial growth for all of the plant pathogens in vitro except S. sclerotiorum. Minimum inhibitory concentrations (MICs) of bEPA were found in the range of 250–500 μg/ml. Also, bEPA had a detrimental effect on spore germination for all the tested plant pathogens. Three plant pathogenic fungi (F. oxysporum, P. capsici and C. capsici) were subjected to an in vivo anti-fungal screening. bEPA at the initial concentration of 3000 μg/ml had a 100% anti-fungal effect against all of the tested plant pathogens. Concentrations of bEPA corresponding to 1500, 500 and 300 μg/ml were applied to the plants and revealed promising anti-fungal effects, supporting bEPA as a potential anti-fungal agent.     

Antifungal activity in vitro of Aloe vera pulp and liquid fraction against plant pathogenic fungi

The leaf pulp of Aloe vera, designated as the gel, and the bitter, yellow liquid fraction have been tested against pathogens (bacteria and fungi) affecting human and plants. However, their activity for fungal control in commercial industrial crops has not been determined. The objectives of this study were to evaluate the inhibitory effect of Aloe pulp and liquid fraction on the mycelial growth of three phytopathogenic fungi and to determine the extract concentrations that can inhibit mycelial development. A. vera leaves were cut from plants grown under greenhouse conditions at the University Antonio Narro, disinfected with sodium hypochlorite, and separated in two groups. In the first group, the pulp was manually scraped out; in the second, a laboratory roll processor was used for the pulp and liquid fraction separation. Both types of extracts were pasteurized. Antifungal activity of pulp and liquid fraction was evaluated on the mycellium development of Rhizoctonia solani, Fusarium oxysporum, and Colletotrichum coccodes that were isolated from a potato crop by the hyphae point and monosporic techniques. The concentrations of the plant extract ranged from 0 to 10⁵ μl l⁻¹. Fungal plugs 0.4 mm in diameter were placed in Petri dishes with a potato–dextrose–agar (PDA) culture media, and treated with various concentrations of pulp or liquid fraction. The cultures were incubated at 24±2 °C and the radial growth of mycelia measured daily for 7 days. The antifungal effect was measured under a totally random design with four replications. The results showed an inhibitory effect of the pulp of A. Vera on F. oxysporum at 10⁴ μl l⁻¹ and over a long period. For the two types of Aloe fractions the activities were similar. Besides the liquid fraction reduced the rate of colony growth at a concentration of 10⁵ μl 1⁻¹ in R. solani, F. oxysporum, and C. coccodes. This is the first report of any Aloe liquid fraction activity against plant pathogenic fungi.     

Antifungal activity in vitro of Flourensia spp. extracts on Alternaria sp., Rhizoctonia solani,and Fusarium oxysporum

There are 32 species of Flourensia genus with 9 native to Mexico. These species contain compounds with potential use for pest control. In this paper, we report the antifungal activities of ethanol extracts from three endemic species in Coahuila state: Flourensia microphylla, Flourensia cernua, and Flourensia retinophylla. Also, preliminary information on the chemical composition of the extracts is included. Antifungal activity was tested against three pathogens attacking commercial crops: Alternaria sp., Rhizoctonia solani, and Fusarium oxysporum. The extracts concentration varied from 10 to 1500 μl l⁻¹. The ANOVA showed highly significant differences (P ≤ 0.01) with the extracts, the doses, and on the interaction extract × dose. Inhibition effect was observed from 10 μl l⁻¹ in all three species. Total inhibition was found only with F. cernua and F. retinophylla at 1000 μl l⁻¹ for R. solani, the three species inhibited the three pathogens at 1500 μl l⁻¹. Infrared analysis showed similar absorption signals for the extracts of the three species although in different concentration. This suggests that similar compounds may be present. The control of these pathogens by natural compounds is interesting both for environmental and economic reasons. The use of semiarid lands plants may improve the socioeconomic level of the people within the region.     

Verification of Glyphosate Resistance,Lepidopteran Resistance and Wide Compatibility of Male Sterile Line E1C4008S in Rice

Weeds and insect pests are two important biotic stresses resulting in yield loss in rice, and wide compatibility is the essential characteristic of breeding inter-subspecific hybrid rice. This study focused on glyphosate resistance, lepidopteran resistance and wide compatibility as well as identification of molecular and some agronomic characteristics of transgenic male sterile line E1 C4008 S. The results indicated that glyphosate resistance gene Epsps# and lepidopteran resistance gene Cry1 ca~# were transferred into japonica wide compatibility male sterile line 4008 S by Agrobacterium-mediated method, and four independent transformation events named E1 C4008 S-1, E1 C4008 S-2, E1 C4008 S-3 and E1 C4008 S-4 were obtained, in which E1 C4008 S-3 and E1 C4008 S-4 were of single copy insertion. The EPSPS protein contents of E1 C4008 S-3 and E1 C4008 S-4 in different organs were significantly different both in descending order of leaf stem root, and ranged from 300.58 to 1410.69 μg/g at the tillering stage. The glyphosate tolerable concentration(dosage) of E1 C4008 S-3 and E1 C4008 S-4 reached at least 16 g/L(54.42 kg/hm2), and the seeds of E1 C4008 S-4 can germinate normally on the medium containing 1 g/L glyphosate. The CRY1 C protein contents of E1 C4008 S-3 and E1 C4008 S-4 in different organs were significantly different both in descending order of leaf stem root, and ranged from 0.62 to 2.43 μg/g at the tillering stage. The larvae mortalities of rice leaf rollers fed on leaves of E1 C4008 S-3 and E1 C4008 S-4 for 5 d were 95.35% and 97.77%, respectively, while the average mortalities of silkworms fed with protein extracts from leaves of E1 C4008 S-3 and E1 C4008 S-4 reached 94.55% and 83.64%, respectively. The results suggested that wide compatibility and evaluated agronomic traits of E1 C4008 S-4 were not significantly changed by insertion of the exogenous genes. Overall, a novel male sterile germplasm with glyphosate resistance, lepidopteran resistance and wide compatibility was verified to be developed in rice.     

Breeding Novel Short Grain Rice for Tropical Region to Combine Important Agronomical Traits,Biotic Stress Resistance and Cooking Quality in Koshihikari Background

Breeding program strategies to develop novel short grain white rice varieties such as japonica (short grain) that introgress biotic stress resistance and high grain quality have been developed using indica rice (Pin Kaset + 4 and Riceberry) for applications in japonica rice (Koshihikari) improvement. Four breeding lines showing promising agronomic performance with short grain and low amylose content (< 20%) were obtained. In addition, sensory testing of these breeding lines showed high scores that similar to Koshihikari. Two promising lines, KP48-1-5 and KP48-1-9, which possessed a combination of four genes resistance to different biotic stresses (Bph3 + TPS + Xa21 + Pi-ta) and four genes for grain quality (GS3 + SSIIa + wx^b + badh2), were developed using marker-assisted selection (MAS) with the pedigree method. The current study clearly illustrated the successful use of MAS in combining resistance to multiple biotic stresses while maintaining a high yield potential and preferred grain quality. Moreover, the results indicated that this breeding program, which includes crossing temperate japonica with indica, can create novel short grain rice varieties adapted to a tropical environment, like the japonica type.     

Appraisal of Biofertilizers in Rice: To Supplement Inorganic Chemical Fertilizer

A field experiment was carried out to evaluate the feasibility of inoculating rice seedlings with biofertilizers (Azospirillum and Trichoderma) in order to reduce the use of chemical inorganic nitrogen (N) fertilizer on rice variety BU Dhan 1. The plant performances were better when 25% less inorganic N was applied with Trichoderma and combined application of Trichoderma and Azospirillum. Plants contained the highest chlorophyll concentrations when they were treated with 75% N + Trichoderma. Considering the yield attributes, 75% N + Trichoderma and 75% N + Trichoderma + Azospirillum performed similar to the control. The grain yield of rice was similar to the recommended dose even with 25% less N application. Application of Trichoderma resulted higher yield, followed by combined application with Azospirillum. Results revealed the greater scope of applying biofertilizer (Trichoderma) to supplement chemical N fertilizer with optimum yield of rice.     

Maximizing yields in rice–groundnut cropping sequence through integrated nutrient management

Integrated use of organic and inorganic fertilizers can improve crop productivity and sustain soil health and fertility. The present research was conducted to study the effects of application of green manures [sesbania (Sesbania aculeate Poiret) and crotalaria (Crotalaria juncea L.)] and farmyard manure on productivity of rice (Oryza sativa L.) and its residual effects on subsequent groundnut (Arachis hypogaea L.) crop. Rice and groundnut crops were grown in sequence during rainy and post-rainy seasons with and without green manure in combination with different fertilizer and spacing treatments under irrigated conditions. The results showed that application of green manures sesbania and crotalaria at 10 t ha⁻¹ to rice compared to no green manure application significantly increased grain yield of rice by 1.6 and 1.1 t ha⁻¹, and pod yields of groundnut crop succeeding rice by 0.25 and 0.16 t ha⁻¹, respectively. There was no significant difference between the application of crotalaria or farmyard manure at 10 t ha⁻¹ on grain yields of rice, but pod yields of subsequent groundnut crop were greater with application of green manure. There was no significant effect of different spacing 20×15,15×15,15×10 cm² (333 000; 444 000; 666 000 plant ha⁻¹, respectively) on grain yield of rice. Pod yields of groundnut were significantly greater with closer spacing 15×15 cm² (444 000 plants ha⁻¹) as compared to spacing of 30×10 cm² (333 000 plants ha⁻¹). Maximum grain of rice was obtained by application of 120:26:37 kg NPK ha⁻¹ in combination with green manures, whereas maximum pod yield of groundnut was obtained by residual effect of green manure applied to rice and application of 30:26:33 kg NPK ha⁻¹ in combination with gypsum applied to groundnut crop.     

Cover Crops as Affecting Soil Chemical and Physical Properties and Development of Upland Rice and Soybean Cultivated in Rotation

Cover crops can provide changes in soil chemical and physical properties, which could allow a sustainable development of soybean and upland rice rotation in Brazilian Cerrado. The objective of this study was to determine the effects of cover crops(cultivated in the offseason) in the soybean-upland rice rotation(cultivated in the summer season) on the soil chemical and physical properties, yield components and grain yield of the cash crops. The experimental design was a randomized block design in factorial scheme 4 × 2 with six replications. Treatments were composed by four cover crops: fallow, millet(Pennisetum glaucum) + Crotalaria ochroleuca, millet + pigeon pea(Cajanus cajans), and millet + pigeon pea + Urochola ruziziensis in the offseason with one or two cycles of cover crops, with rice(Oryza sativa)or soybean(Glycine max) in the summer season. Cover crops alone provided no changes in soil chemical properties. However, the rotation cover crops/cash crops/cover crops/cash crops reduced p H, Al and H + Al and increased Ca, Mg, K and Fe contents in the soil. The cover crops millet + pigeon pea and millet + pigeon pea + U. ruziziensis improved soil physical properties in relation to fallow,especially in the 0–0.10 m soil layer. In spite of the improvement of the soil physical properties after two years of rotation with cover crops and cash crops, the soil physical quality was still below the recommended level, showing values of macroporosity, S index and soil aeration capacity lower than 0.10 m3/m3, 0.035 and 0.34, respectively. Upland rice production was higher under mixtures of cover crops than under fallow, mainly because of soil physical changes done by these mixtures of cover crops.Soybean grain yield was similar under all cover crops tested, but was higher after the rotation cover crops/upland rice/cover crops than after only one cycle of cover crops.     

Prediction and Expression Analysis of miRNAs Associated with Heat Stress in Oryza sativa简

Computational prediction of potential microRNAs (miRNAs) and their target genes was performed to identify the miRNAs and genes associated with temperature response in rice. The data of temperature-responsive miRNAs of Arabidopsis, and miRNAs and the whole genome data of rice were used to predict potential miRNAs in Oryza sativa involved in temperature response. A total of 55 miRNAs were common in both the species, and 27 miRNAs were predicted at the first time in rice. Target genes were searched for these 27 miRNAs in rice genome following stringent criteria. Real time PCR based on expression analysis of nine miRNAs showed that majority of the miRNAs were down regulated under heat stress for rice cultivar Nagina 22. Furthermore, miR169, miR1884 and miR160 showed differential expression in root and shoot tissues of rice. Identification and expression studies of miRNAs during heat stress will advance the understanding of gene regulation under stress in rice.     

The efficiency of nitrogen fertiliser for rice in Bangladeshi farmers’ fields

Bangladesh is currently self sufficient in rice (Oryza sativa L.), which accounts for approximately 80% of the total cropped area, and 70% of the cost of crop production. However, farmers are increasingly concerned about the perceived decline in productivity, expressed as the return on fertiliser inputs. Agronomic efficiency is a measure of the increase in grain yield achieved per unit of fertiliser input that can provide a way to quantify the observation of farmers. This study indicates that the yields achieved where only P and K fertiliser were applied ranged from 3–5 t ha⁻¹, indicating good soil fertility, particular in terms of soil N supply (37–112 kg N ha⁻¹). However, at recommended rates and at rates used by farmers, the yield response to application of fertiliser N was low. Data shows that grain yields were significantly correlated in both years (R² = 0.77 and R² = 0.67) with plant uptake in nitrogen. The internal nitrogen use efficiency seems to confirm that sink formation was limited by factors other than nitrogen. Low agronomic efficiency (5–19 kg grain kg⁻¹ N) was caused by poor internal efficiency (45–73 kg grain kg⁻¹ N), rather than low supply of soil N or loss of fertiliser N. Thus, often the applications of large amounts of N fertiliser (39–175 kg N ha⁻¹) by farmers to increase yields of high yielding variety Boro rice were not justified agronomically and ecologically. A rate of 39 kg N ha⁻¹ is very low, hardly an environmental threat. No one single factor could be identified to explain the low internal efficiency. Therefore, it is concluded that the data presented tend to confirm the indication that yields are limited by a factor other than nitrogen, which could be crop establishment, plant density, water or pest management, micro-nutrients deficiency, poor seed and transplanted seedling quality, varieties and low radiation.     

A comparison of the lipid and fatty acid profiles from the kernels of the fruit (nuts) of Ximenia caffra and Ricinodendron rautanenii from Zimbabwe

The lipid profile of nuts from Ximenia caffra and Ricinodendron rautanenii was determined and compared. Although the total oil content of X. caffra and R. rautanenii nuts was similar (47.6 ± 7.5% versus 53.3 ± 13.7%), the fatty acid profiles differed significantly. X. caffra had a higher content (p < 0.05) of saturated fatty acids than R. rautanenii (20.19 ± 1.07% versus 13.87 ± 3.68%) and contained C22:0 and C24:0 which were lacking in R. rautanenii. Total monounsaturated fatty acids were higher in X. caffra than R. rautanenii (71.48 ± 0.99% versus 36.66 ± 1.95%). Oleic acid (C18:1n9) was the major monounsaturated fatty acid (MUFA) in X. caffra whereas erucic acid (C22:1n9), the major MUFA in R. rautanenii, was undetectable in X. caffra. R. rautanenii had a greater polyunsaturated fatty acid content than X. caffra which contained C18:3n3 (α-linolenic acid) and nervonic acid (24:1n9). X. caffra is potentially an important source of essential fatty acids.     

Productivity and residual effects of legumes in rice-based cropping systems in a warm-temperate environment: II. Residual effects on rice

Field experiments were conducted over the period 1994–1996 to investigate the residual effect on rice of a wide range of temperate legume species, grown during the preceding winter season in a warm-temperate environment in Nepal. The incorporation of large quantities of above- and below-ground legume biomass (roots and foliage) resulted in substantial residual effects on the subsequent upland rice crop. The effects on rice yield in terms of percent increase over the control were more pronounced at a site of comparatively lower inherent soil fertility where grain yields of rice grown after high yielding legume crops such as bitter lupin (Lupinus mutabilis) and Persian clover (Trifolium resupinatum) were almost twice as high (up to 7.6 t DM ha⁻¹) than those of the control treatment (rice after wheat). In contrast, the residual effects of below-ground biomass (roots, nodules) on the following rice crop were not very pronounced and resulted in similar residual effects as rice, after fallow. Although up to 480 kg ha⁻¹ of legume foliage N had been applied to rice, only a small proportion of the legume N (up to 70 kg N ha⁻¹) was recovered by the following rice crop. Total rice dry matter yields were highly correlated with the amount of legume N applied, yet other factors such as residue quality and residue management practices appeared to also affect the magnitude of the residual effects. Future work should aim at investigating the effect of residue quality as affected by legume species and residue management practice on decomposition and N mineralisation processes in rice soils.     

Integrating rice and wheat productivity trends using the SAS mixed-procedure and meta-analysis

Long-term trends of crop yields have been used as a means to evaluate the sustainability of intensive agriculture. Previous studies have measured yield trends from long-term rice–rice and rice–wheat experiments in different sites from the slopes of individual site regressions of yield over time. The statistical significance of each site regression was determined but not that of the aggregate trend, which could give an indication of the magnitude and significance of global yield change.The random regression coefficient analysis (RRCA) and meta-analysis were used in this study to analyze the aggregate yield trend from several long-term experiments (LTE) across the Indo-Gangetic Plains (IGP) and outside the IGP. Both methods show that there has been a significant (p < 0.05) declining trend in rice yield in rice–wheat LTEs in South Asia including China with the recommended rates of nutrients, but that there has been no significant change in wheat and system (rice + wheat) yields. There was no significant year × region (IGP versus non-IGP) interaction in rice and wheat yields. However, RRCA showed that the average yield trend was significantly negative (−41.0 kg ha⁻¹ yr⁻¹) only in the IGP. In the rice–rice LTEs, there was a significant year × site (IRRI versus non-IRRI sites) interaction during the dry season but not the wet season. Rice yields declined throughout Asia in the wet season. The average system (dry + wet season rice) yield trends were significantly negative in both IRRI and non-IRRI sites (−170.1 and −52.8 kg ha⁻¹ yr⁻¹, respectively) but the magnitude of yield decline was significantly greater in the IRRI sites than in the non-IRRI sites.Rice in the rice–wheat LTEs showed a significantly positive yield trend with the addition of farmyard manure (FYM) but the initial yield was generally lower with FYM than without FYM. After 15 years, yield increase due to FYM was not evident in most of the LTE.     

Intra-specific competition in maize: early establishment of hierarchies among plants affects final kernel set

Reduced plant biomass and increased plant-to-plant variability are expected responses to crowding in monocultures, but the underlying processes that control the onset of interplant interference and the establishment of hierarchies among plants within a stand are poorly understood. We tested the hypothesis that early determined plant types (i.e. dominant and dominated individuals) are the cause of the large variability in final kernel number per plant (KNP) usually observed at low values of plant growth rate (PGR) around silking in maize (Zea mays L.). Two hybrids (DK696 and Exp980) of contrasting response to crowding were cropped at different stand densities (6, 9 and 12 plants m⁻²), row spacings (0.35 and 0.70 m), and water regimes (rainfed and irrigated) during 1999/2000 and 2001/2002 in Argentina. The onset of interplant competition started very early during the cycle, and significant differences (P<0.05) in estimated plant biomass between stand densities were detected as soon as V_4–6 (DK696) and V_6–7 (Exp980). Plant population and row spacing treatments did not modify the onset of the hierarchical growth among plants, but did affect (P<0.02–0.08) the dynamic of the process. For both hybrids, the rate of change in relative growth between plant types was larger at 9 and 12 plants m⁻² (ca. 0.12 g/g per 100 °C day) than at 6 plants m⁻² (ca. 0.07 g/g per 100 °C day). For all treatments, the largest difference in estimated shoot biomass between plant types took place between 350 (V₇) and 750 °C day (V₁₃) from sowing, and remained constant from V₁₃ onwards. Dominant plants always had more kernels per plant (P<0.05) than the dominated ones, but differences between plant types in PGR around silking were significant (P<0.05) only at 12 plants m⁻². Our research confirmed the significant (P<0.01) curvilinear response of KNP to PGR around silking, but also determined a differential response between plant types: the mean of residual values were significantly (P<0.01) larger for dominant than for dominated individuals. Estimated ear biomass at the onset of active kernel growth (R₃) reflected the variation in KNP (r²≥0.62), and was significantly (P<0.01) related to estimated plant biomass at the start of active ear growth (ca. V₁₃). This response suggested that the physiological state of each plant at the beginning of the critical period had conditioned its reproductive fate. This early effect of plant type on final KNP seemed to be exerted through current assimilate partitioning during the critical period.     

Long-term effects of inorganic and organic fertilizer sources on yield and nutrient accumulation of lowland rice

Deceleration in rice (Oryza sativa L.) yield over time under fixed management conditions is a concern for countries like Bangladesh, where rice is the primary source of calories for the human population. Field experiments were conducted from 1990 to 1999 on a Chhiata clay loam soil (Hyperthermic Vertic Endoaquept) in Bangladesh, to determine the effect of different doses of chemical fertilizers alone or in combination with cow dung (CD) and rice husk ash (ash) on yield of lowland rice. Two rice crops—dry season rice (December–May) and wet season rice (July–November) were grown in each year. Six treatments—absolute control (T₁), one-third of recommended fertilizer doses (T₂), two-thirds of recommended fertilizer doses (T₃), full doses of recommended fertilizers (T₄), T₂+5 t CD and 2.5 t ash ha⁻¹ (T₅) and T₃+5 t CD and 2.5 t ash ha⁻¹ (T₆) were compared. The CD and ash were applied on dry season rice only. The 10-year mean grain yield of rice with T₁ was 5.33 t ha⁻¹ per year, while the yield with T₂ was 6.86 t ha⁻¹ per year. Increased fertilizer doses with T₃ increased the grain yield to 8.07 t ha⁻¹ per year, while the application of recommended chemical fertilizer doses (T₄) gave 8.87 t ha⁻¹ per year. The application of CD and ash (T₅ and T₆) increased rice yield by about 1 t ha⁻¹ per year over that obtained with chemical fertilizer alone (T₂ and T₃, respectively). Over 10 years, the grain yield trend with the control plots was negative, but not significantly, both in the dry and wet seasons. Under T₃ through T₆, the yield trend was significantly positive in the dry season, but no significant trend was observed in the wet season. The treatments, which showed positive yield trend, also showed positive total P uptake trend. Positive yield trends were attributed to the increasing P supplying power of the soil.     

Characterization and Evaluation of OsLCT1 and OsNramp5 Mutants Generated Through CRISPR/Cas9-Mediated Mutagenesis for Breeding Low Cd Rice

To explore how rice(Oryza sativa L.) can be safely produced in Cd-polluted soil, OsLCT1 and OsNramp5 mutant lines were generated by CRISPR/Cas9-mediated mutagenesis. One of OsLCT1 mutant(lct1×1) and two of OsNramp5 mutants(nramp5×7 and nramp5×9) were evaluated for grain Cd accumulation and agronomic performances. In paddy field soil containing approximately 0.9 mg/kg Cd, lct1×1 grains contained approximately 40%(0.17 mg/kg) of the Cd concentration of the wild type parental line, less than the China National Food Safety Standard(0.20 mg/kg). Both OsNramp5 mutants showed low grain Cd accumulation(< 0.06 mg/kg) in the paddy(approximately 0.9 mg/kg Cd) or in pots in soil spiked with 2 mg/kg Cd. However, only nramp5×7 showed normal growth and yield, whereas the growth of nramp5×9 was severely impaired. The study showed that lct1×1 could be used to produce rice grains safe for human consumption in lightly contaminated paddy soils and nramp5×7 used in soils contaminated by much higher levels of Cd.     

Fine Mapping of QTLs for Stigma Exsertion Rate from Oryza glaberrima by Chromosome Segment Substitution

Stigma exsertion is an important trait for outcrossing ability in rice. Stigma exsertion rate (SER) of male sterile lines (MSLs) is a key factor affecting F₁-seed production in hybrid rice. In this study, seven QTLs for SER were detected on five chromosomes using a set of single-segment substitution lines (SSSLs) derived from O. glaberrima. Three of the QTLs were mapped in the estimated intervals of 92.5–333.0 kb. qSER-5 was located in a substitution segment of 92.5 kb. qSER-1b and qSER-8b were respectively limited to 333.0 kb and 107.5 kb by secondary substitution mapping. qSER-1b and qSER-3 had bigger additive effects of 11.5% and 11.9%, respectively, while the other five QTLs had smaller additive effects from 5.7% to 8.6%. Open reading frames were identified in the regions of qSER-5 and qSER-8b in O. sativa and O. glaberrima genomes. Fine mapping of the QTLs laid a foundation for the cloning of genes, and QTLs for SER will be used to develop MSLs with strong ability of outcrossing.