Performance of Five Early Indica Rice Varieties under Direct Seeding in Jiangbei District

Comparative experiments were conducted on five conventional indica rice varieties,including Yongxian 15,Zhongzao 39,Shunda 135,Zhongzu 143,and Zhongzu 18.Under direct seeding conditions,their growth period and yield performance were investigated.The results showed that the yield of Zhonggang 143 and Shunda 135 was higher than that of Yongxian 15(CK),and Zhonggang 143 had the highest yield of 7.494 t/hm 2,followed by Shunda 135(7.467 t/hm 2);Yongxian 18(CK)has a yield of 7.326 t/hm 2.Taking into account factors such as field growth,yield,and growth period,Zhongzu 143 and Shunda 135 can be further promoted to optimize the early rice variety structure in Jiangbei District.

Developmental Genetic Analysis of Brown Rice Weight Under Different Environmental Conditions in indica Rice

Analysis of genetic main effects and genotype x environment (GE) interaction effects for brown rice weight (BRW) at four different filling stages in indica lice (Oryza sativa L.) was conducted for two-year experimental data by using developmental genetic models and corresponding statistical approaches for quantitative traits of seeds in cereal crops. It was indicated that the genetic main effects and their GE interaction effects of triploid endosperm, cytoplasmic and diploid maternal plant genes were important for BRW at different filling stages of rice, especially for endosperm or maternal additive main effects and their additive interaction effects. Because of the higher additive effects and additive interaction effects for BRW at different filling stages, the better improving effects for this trait could be expected by selection in rice breeding. The results of conditional genetic variance components showed that the new expression of quantitative genes in endosperm and maternal plant for BRW was mostly found at all different filling stages of rice. The gene expression, however, was most active at the early filling stages especially for the first (1-7 d) and the second filling stages (8-14 d after flowering). The phenomena that some genes were spasmodically expressible among filling stages of rice were detected for some genetic effects especially for net cytoplasmic main effects or its interaction effects and net dominance main effects. Predicted genetic effects at different filling stages of rice showed that some parents such as V20 and Zuo 5 were better than others for improving the BRW.

Analysis of the Conditional Correlations from Different Genetic Systems Between the Protein Content and the Appearance Quality Traits of Indica Rice

A factorial mating design in two environments was conducted using 7 cytoplasmic male sterile lines （A） and 5 restorer lines （R） along with their F1 （A × R） and F2 populations. The unconditional and conditional analyses of genetic models and the corresponding statistic methods, including endospermic, cytoplasmic, and maternal plant genetic systems, were used to analyze the genetic relationships between protein content （PC） and the appearance quality traits of indica rice （Oryza sativa L.）. The results from unconditional analysis indicated that PC was significantly correlated with the appearance quality traits of rice, except for the brown rice thickness （BRT）. Only the genetic covariance between PC and the brown rice width （BRW） was positively correlative, whereas all the other pairwise traits were negatively correlative. The results from conditional analysis revealed that the weight of brown rice （WBR） or the amylose content （AC） could significantly affect the relationships between PC and the appearance quality traits of indica rice. The conditional analysis showed that WBR might negatively affect the relationships between PC and the brown rice length （BRL）, BRW, or BRT through the geuotype x environmental （GE） interaction effects, but positively affected the relationships between PC and the ratio of brown rice length to width （RLW） or the ratio of brown rice length to thickness （RLT）. The amylase content could positively affect the relationships between PC and BRL, RLW, RLT through the cytoplasmic effects and maternal additive effects, but negatively affected the relationships between PC and BRW.

Development of Isogenic Lines of Morphological Markers in Indica Rice

A total of 28 accessions of rice (Oryza sativa L.) germplasms were collected from International Rice Research Institute (IRRI), Philippine and Gene Pool of China National Rice Research Institute (CNRRI). These germplasms were attributed as a Donor Gene Pool, consisting of 27 diverse morphological marker lines with each having a unique valuable trait (gene), and the recurrent parent-Zhefu 802, an early-season indica cultivar in south of China in the 1980s and 1990s. A set of 27 isogenic lines with 2-3 markers on every chromosome were obtained after more than 10 successive backcrosses. Moreover, 15 accessions of double-marker isogenic lines were developed by further pyramiding markers on the same chromosome. Key agronomic traits such as heading date, plant height, tillering number and panicle length in this set of lines were similar to that of Zhefu 802 except the marker traits.

Analysis of Genetic Variation and Population Structure of Starch Synthesis-related Genes in Indica Rice Cultivars

In this study, 34 molecular markers of starch synthesis-related genes were used to evaluate the genetic variation and population structure of 87 indica rice cultivars from different countries and regions. The results showed that a total of 80 alleles were amplified using 34 primer pairs, with an average of 2.5 alleles per locus. The allele number varied from 2 to 6 among various cultivars. Shannon＇s diversity index of molecular markers varied from 0.303 to 0.796, with an average of 0.539. Polymorphism information content （PIC） varied from 0.084 to 0.658, with an average of 0.295. The genetic similarity coefficients of 87 indica rice cultivars ranged from 0.265 to 0.990, indicating significant genetic differences of starch synthesis-related genes among different cultivars, but the variation frequency of alleles varied among different cultivars. Population structure analysis showed that these 87 indica rice cultivars were divided into three categories. Genetic differences were small within the same category but great among different categories. Moreover, indica rice cultivars with simple genetic components accounted for 39.1% and those with complex genetic background accounted for 60.9%. This study may not only provide theoretical basis for genetic improvement of rice starch quality, but also lay a solid foundation for subsequent association analysis of rice quality-related traits.

Synthesis and paste properties of octenyl succinic anhydride modified early Indica rice starch

Octenyl succinic anhydride （OSA） modified early Indica rice starch was prepared in aqueous slurry systems using response surface methodology. The paste properties of the OSA starch were also investigated. Results indicated that the suitable parameters for the preparation of OSA starch from early Indica rice starch were as follows： reaction period 4 h, reaction temperature 33.4℃, pH of reaction system 8.4, concentration of starch slurry 36.8% （in proportion to water, w/w）, amount of OSA 3% （in proportion to starch, w/w）. The degree of substitution was 0.0188 and the reaction efficiency was 81.0%. The results of paste properties showed that with increased OSA modification, the starch derivatives had higher paste clarity, decreased retrogradation and better freeze-thaw stability.

Difference of Nitrogen Uptake and Use Efficiency in Conventional Indica Rice Cultivars with Different Nitrogen Use Efficiency for Grain Output

[Objective] The study aimed to confirm difference of nitrogen uptake and used efficiency with different nitrogen use efficiency for grain output (NUEg) types of indica rice.[Method] 88 and 122 conventional indica rice cultivars were solution-cultured in 2001 and 2002, respectively. Dry matter weight (including root system, culm and sheath, leaves, panicle), nitrogen content of different organs, yield and its components were measured. The tested rice cultivars were classified into 6 types (i.e. A, B, C, D, E and F, A was the lowest, and F was the highest) based on their NUEg level by the MinSSw method.[Result](1)Difference of NUEg of the cultivars used in this study were very large; (2) No significant difference of N content at heading stage was observed among different NUEg types of indica rice. In the cultivars with higher NUEg, however, N content in leaf, stem-sheath and entire rice plant were lower at mature stage. (3)Cultivars with higher NUEg were characterized with lower N uptake before heading and at mature stage; (4) Cultivars with higher NUEg were characterized with higher N use efficiency in biomass production and harvest index. [Conclusion] The cultivars with higher NUEg showed lower N uptake and N content, while nitrogen use efficiency was higher.

Quantitative Trait Loci for Grain Chalkiness and Endosperm Transparency Detected in Three Recombinant Inbred Line Populations of Indica Rice

Quantitative trait loci（QTL） for percentage of chalky grain,degree of chalkiness,and endosperm transparency were detected using 3 recombinant inbred line populations derived from crosses between parental lines of commercial three-line hybrids of indica rice.Two of the populations showed great variations on heading date,and the other had a short range of heading date variation.A total of 40 QTLs were detected and fell into 15 regions of 10 chromosomes,of which 5 regions were detected for 1 or more same traits over different populations,2 were detected for different traits in different populations,3 were detected for 2 or all the 3 traits in a single population,and 5 were detected for a single trait in a single population.Most of these QTLs have been reported previously,but a region located on the long arm of chromosome 10 showing significant effects in all the 3 populations has not been reported before.It was shown that a number of gene cloned,including the Wx and Alk for the physiochemical property of rice grain,and GW2,GS3 and GW5 for grain weight and grain size,could have played important roles for the genetic control of grain chalkiness in rice,but there are many more QTLs exerting stable effects for rice chalkiness over different genetic backgrounds.It is worth paying more attentions to these regions which harbor QTL such as the qPCG5.2/qDC5.2/qET5.2 and qPCG10/qDC10/qET10 detected in our study.Our results also showed that the use of segregating populations having high-uniform heading date could greatly increase the efficiency of the identification of QTL responsible for traits that are subjected to great environmental influence.

Natural Variation of Leaf Thickness and Its Association to Yield Traits in indica Rice

Leaf thickness is an important morphological trait in rice. Its association to the yield potential, as of now has not been documented because of the shortage of the equipment which could conveniently measure the leaf thickness in rice. In this study, the thickness of top three leaves of 208 cultivars had been determined by a nondestructive rice leaf thickness instrument for the research of the natural variation of leaves thickness and its association to yield traits in indica rice. The results showed that the flag leaf was the thickest, and the 2nd leaf was thicker than the 3rd leaf. Analysis of variance indicated the existence of wide genetic diversity of leaf thickness among the investigated indica rice genotypes. The tight correlation among the thicknesses of the top three leaves means that the leaf thickness traits share one genetic control system. Leaf thickness had a significant positive correlation with leaf length and a positive correlation with leaf width, indicated that thicker leaf was beneficial to increasing the single leaf area. The results of correlation analysis revealed that thicker leaf should be profitable to the leaf erection, higher numbers of grains per panicle and higher grains weight per panicle. However, the significantly negative correlation between leaf thickness and number of panicles per plant counteracted the profitability from increased grains weight per panicle, so that the correlations of the thicknesses of the top three leaves to yield and biomass were positive but not significantly. It has made great progress in the genetic improvement of leaves thickness in inbred indica rice breeding in Guangdong Province, China, since the 1990s.

Effects of different mechanical direct seeding methods on grain yield and lodging resistance of early indica rice in South China

Direct seeding of rice has become a main planting method due to the low labor input and high economic benefit in South China.Dry direct seeding(DDS)has been widely used for single-season rice planting establishment.However,few studies have examined the performance of early-season indica rice under mechanical dry direct seeding.A two-year field experiment was conducted with two indica rice cultivars(i.e.,Zhongjiazao 17 and Zhuliangyou 819)to study lodging characteristics and grain yield formation under DDS,flooded direct seeding(FDS)and wet direct seeding(WDS)patterns.The results showed that the annual grain yield in DDS was higher by 14.42–26.34%for cultivar ZLY819 and 6.64–24.58%for cultivar ZJZ17 than in WDS and FDS,respectively,and these increases were mainly attributed to the improvement of the panicles.The DDS pattern significantly increased the seedling emergence rate of early indica rice cultivars,and increased total dry weight and crop growth rate.Meanwhile,shorter basal internodes,better stem diameter and stem wall thickness and lower lodging index were found in DDS in contrast to FDS and WDS.In particular,DDS improved the stem lodging resistance.Our results suggested that the appropriate direct seeding method was beneficial for improving the grain yield and lodging resistance of early indica rice.

Response of grain-filling rate and grain quality of mid-season indica rice to nitrogen application

Nitrogen is one of the important factors for high yield of rice.Apart from high yield,high quality has become the current urgent demand for rice production.Grain-filling stage is crucial for rice yield and quality formation.However,the effect of nitrogen on grain-filling characteristics and the relationship of grain-filling characteristics and rice quality of mid-season indica rice were still unclear.A field experiment was carried out to ascertain the critical grain-filling characteristics that contribute to rice milling quality,appearance quality and cooking and eating quality under nitrogen applications.The results showed that nitrogen applications prolonged the duration of superior and inferior grain filling.The mean grainfilling rate(G_(mean))and the maximum grain-filling rate(G_(max))of the inferior grains were positively correlated with chalky kernel rate,chalkiness,and amylose content.The time reaching the maximum grain-filling rate(T_(max)G)of the inferior grains was positively correlated with brown rice rate,milled rice rate,and head milled rice rate.Chalky kernel rate and chalkiness were negatively correlated with peak paste viscosity and breakdown viscosity.Less amylose content and more crude protein content were detected in nitrogen application of Liangyoupei 9 and Y Liangyou 2 both in 2016 and 2017.According to the correlation analysis,better cooking and eating quality of Y Liangyou 2 which had less amylose content might result from its higher G_(max)and G_(mean)of inferior grain than that of Liangyoupei 9 in the treatments of nitrogen application.These results indicated that the prolonging grain-filling duration and increasing grain weight at the maximum grain-filling rate of inferior grains contributed to the improvement of milling quality,appearance quality,and cooking and eating quality of mid-season indica rice under appropriate nitrogen applications.

Effects of increasing panicle-stage N on yield and N use efficiency of indica rice and its relationship with soil fertility

Because of the higher nitrogen(N)recovery efficiency(NRE)of panicle-stage fertilization compared with basal and tillering fertilization,increasing the proportion of N topdressing at the booting stage(panicleN)is recommended and commonly practiced in parts of China.To investigate the effects of increasing panicle-N on grain yield and N use efficiency(NUE)and the relationships of the increase and the rice cultivar and soil fertility status,we increased the percentage of panicle-N from 20%to 40%by correspondingly reducing the N amount applied only at the tillering stage in both high-and low-fertility blue clayey paddy fields in 2018 and 2019.Four indica cultivars with diverse panicle types were used,and their grain yield,dry matter accumulation,and NUE were compared.In high-fertility soil,increasing topdressing panicle-N from 20%to 40%reduced tillering ability and reduced the effective panicle numbers of the multi-and medium-panicle cultivars Huanghuazhan(HHZ),C Liangyouhuazhan(CHZ),and Tianyouhuazhan(THZ).These cultivars gave the greatest yield when 30%of N was supplied as panicle fertilizer,whereas the yield,NRE,N agronomic efficiency(NAE),and nitrogen physiological efficiency(NPE)of the heavy-panicle inbred cultivar Yangdao 6(YD6)continued to increase,resulting in improved dry matter accumulation and grain filling in the late growth stage.The yield,NAE,NRE,and NPE of YD6 peaked when the panicle-N constituted 40%.While in low-fertility soil,the multipanicle cultivar HHZ showed the greatest yield when 30%of fertilizer-N was applied once at the panicle initiation(PI)stage,while the medium-panicle cultivar CHZ showed the greatest yield when the panicle-N percentage was 40%.Our results suggest that the percentage of panicle-N fertilizer should not exceed 30%for multipanicle cultivars,while can be appropriately increased to 40%for heavy-panicle indica cultivars.The effect of increasing topdressing panicle-N on the yield of medium-panicle cultivars was related to soil fertility.The optimum panicle-N percentage was 30%in the high-fertility soil and 40%in the low-fertility soil.

Optimization of Electroporation Parameters for Immature Embryos of indica Rice (Oryza sativa)

To obtain a suitable condition for electroporation transformation in indica rice, the 10-day-old immature embryos were selected for optimization experiments. The results showed that one pulse at 850 V/cm, 950μF capacitance, 200 μL electroporation buffer with 70 mmol/L sodium glutamate, 100 μg/mL plasmid, 50μg/mL carrier DNA, 20 embryos per cuvette, 0℃ treatment and CC medium were the best parameters, which not only improved the transformation efficiency to 30.89%, but also ameliorated the embryo survival ratio to 95.92%. To further verify the practicability of this condition, the embryos from another indica rice variety and a rice type Ⅱ metallothionein-like gene （OsMT2bL） promotec：mgfp5：：gusA construct were tested, and specific GUS expression on the embryos was visualized by histochemical staining. The results showed that the GUS expression on the embryos activated by the OsMT2bL promoter was mainly concentrated on the apical point of the plumule whereas the expression driven by CaMV35S promoter was distributed on nearly all areas of the electroporated tissues. These results indicated that the optimized embryo electroporation conditions could be used not only in genetic transformation of indica rice but also in assay of gene regulation on embryos.

Genetic Analysis and QTL Mapping of Mature Seed Culturability in Indica Rice

Genetic segregation analysis for mature seed culturability was conducted using recombinant inbred lines derived from a cross of indica rice, Yangdao 6 and Pei＇ai 64s. Three indices of seed culturability, the frequency of callus induction, the frequency of brown callus and the increase of callus weight were investigated. A combined genetic map constructed with simple sequence repeat （SSR）, sequence tag site （STS）, cleaved amplified polymorphic sequences （CAPS） and single nucleotide polymorphism （SNP） markers covered a total distance of 1 732.5 cM, averaging approximately 12 cM between two neighboring loci. Three QTLs on chromosomes 7, 7 and 10 were detected for the frequency of callus induction; three QTLs on chromosomes 6, 7 and 9 were detected for the frequency of brown callus; and two QTLs on chromosomes 5 and 7 were detected for the increase of callus weight. Common QTLs mapped at the interval flanking RM5481 and RM6835 on chromosome 7 were identified to be involved in the frequency of callus induction and the frequency of brown callus, explaining 7.29% and 12.52% of phenotypic variation, respectively. A total of 14 epistatic effects were detected for the three indices of mature seed culturability. ~

Effect of Temperature on Rice Starch Biosynthesis Metabolism at Grain-Filling Stage of Early Indica Rice

The sucrose content, starch content and the ratio of amylose to total starch and the activities of six key enzymes, sucrose synthase(SS) , soluble starch synthesis enzyme(SSS) , ADPG pyrophosphorylase (ADPG-Ppase) , granule-boundstarch synthase (GBSS), starch branching enzyme (SBE) and debranching enzyme (DBE) , which involved in starch synthesis metabolism of developing rice grains and effect of temperature on their activities were analyzed by using two early indica varieties Zhefu49 and Jiazao935 in phyto-trons, the daily average temperature was 22 and 32℃, respectively. Results showed that the sucrose content, the starch content, and the activities of enzymes such as SS, ADPG-Ppase, SSS, SBE, and DBE were generally higher under the high temperature treatment (32℃)at the beginning stage as compared with that under low temperature treatment (22℃). However, at the middle and late stages, there were great differences in the change trend of some enzyme activities, and the association with the sucrose content and starch content was very complicated. It could be found that the activities of SS and SSS under high temperature were always lower than that under low temperature, which was closely related to the variation of the sucrose content and starch-accumulating rate. Moreover, compared with ADPG-Ppase, the SS and SSS activities were more sensitive to temperature, indicating that SS and SSS were controlled by enzyme through affecting starch accumulating rate. Difference of the amylose/total starch ratio in developing grains between 32 and 22℃was significant and the lower amylose/total starch ratio was always lower in 32℃, which was closely related to the lower activity level of GBSS. Therefore, it could be concluded that effect of temperature on amylose /total starch was more attributable to GBSS rather than to SS, SSS, ADPG-Ppase, SBE and DBE.

Infection of Early and Young Callus Tissues of Indica Rice BPT5204 Enhances Regeneration and Transformation Efficiency

A rapid and reproducible method to develop transgenic plants with enhanced transformation efficiency using Agrobacterium has been developed for the elite indica rice variety BPT 5204. Different rice calli aged from 3 to 30 d were co-cultivated with pre-incubated Agrobacterium suspension culture （LBA4404： pSB1, pCAMBIA1301） and incubated in dark for 3 d. Based on the transient GUS gene expression analysis, 6-day-old young calli showed high transformation frequency followed by 21-day-old ones. Thus, both 6-and 21-day-old calli were used for assessing the stable transformation efficiency. It was observed that the 6-day-old young transformed calli showed about 2-fold higher regeneration frequency when compared with 21-day-old calli. The transformation efficiency was enhanced for young calli to 5.9% compared with 0.8% of the 21-day-old calli. Molecular and genetic analysis of transgenic plants （To） revealed the presence of 1-2 copies of T-DNA integration in transformants and it follows Mendalian ratio in T1 transgenic plants. From the present study, it was concluded that the development of transgenic rice plants in less duration with high regeneration and transformation efficiency was achieved in BPT 5204 by using 6-day-old young calli as explants.

Development and Characterization of Elite Doubled Haploid Lines from Two Indica Rice Hybrids

Despite significant yield advantage over inbred rice, the adoption rate of hybrid rice in India is very low due to the high seed cost and poor quality of the produce. To alleviate the problem, we initiated a doubled haploid （DH） breeding approach to develop new lines from two elite indica rice hybrids （CRHR5 and CRHR7） through rapid fixation of homozygosity in the recombinants. In vitro culture of the rice anthers resulted in 243 and 186 fertile DH lines of CRHR5 and CRHR7, respectively. Flow cytometric and pollen fertility analyses confirmed the DH ploidy status of the regenerations. Morpho-agronomic evaluation revealed 100% uniformity and stability for all the characters in the DH lines of both hybrids. Nineteen promising DH lines of each hybrid were advanced to A2 generation for yield evaluation. The yield levels of the DH lines ranged from 5 097-6 965 kg/hm^2 for CRHR5 and 5 141-7 235 kg/hm^2 for CRHR7, which were at par or higher than the parental hybrids. Physico-chemical characterization and cooking quality analyses revealed significant and acceptable values for grain length and width, alkali spreading value, amylose content and water uptake ratio of the selected DH lines. Three DH lines, CR5-10, CR5-49, CR5-61 from CRHR5, and four DH lines, CR7-5, CR7-7, CR7-12 and CR7-52 from CRHR7, showed significant grain yield and quality characteristics and have been recommended for multi-location trials for subsequent release as new indica doubled haploid rice varieties.

Studies of Improving the Frequency of Indica Rice Transformation by Biolistic Bombardment

In order to improve the frequency of indica rice transformation by biolistic bombardment, suitable culture conditions for embryonic calli,an optimal selection scheme for resistant calli and seedlings, and optimum bombardment parameters a investigated by using 14 commercially important indica rice cultivars. The main results show that the CC medium with 36g/L mannitol is a scheme subculture medium in which the browning of indica rice calli can be mitigated significantly; The concentration of 30～40mg/L Hyg or 150～200mg/L G418 or 10～20 mg/L Basta is suitable for selection of resistant calli; The transformation parameters of 100μg gold powder absorbing 0.2μg DNA per shot and 900 psi helium pressure and 6 cm bombardment distance and bombarded twice for each plate give the best result; Keeping the target calli on osmotic medium containing 60g/L mannitol from 12 ～24h before bombardment to 24～48h after it can increase the efficiencies of transformation . Furthermore, some transgenic indica rice plants are obtained using this optimized transformation system.

Analysis of genetic effects and correlations for nutrient quality traits of indica rice in different environments

Understanding the variation for the expressionof genes in different environments is one of themajor goals in qualitative genetics. In this pa-per, the genetic models for quantitative traitsof endosperm in cereal crops were used to eval-uate the seed, cytoplasm and maternal geneticeffects as well as the genotype × environment(GE) interaction effects, and to predict thebreeding value of parents and genotypic corre-lation for nutrient quality traits of indica rice (Oryza sativa L.).

Caryopsis Development and Main Quality Characteristics in Different indica Rice Varieties

A comparison on the caryopsis development, rice quality characteristics and the size, shape, structure of the endosperm amyloplasts （starch granules） between two indica rice varieties was made, The main quality traits in Yangdao 6 were better than those in Xiangzaoxian 33; In the early period after fertilization the dry matter in Yangdao 6 was accumulated more slowly than that in Xiangzaoxian 33 but faster in the later period, and the starch was accumulated strongly in the later period; There were two kinds of amyloplasts： single and compound amyloplasts, being 4.4 um and 9.5 um in diameter on average with the range of 2.4-8.0 um and 5.7-19.5 um, respectively. In the case of Xiangzaoxian 33, most of the single amyloplasts were elliptic or round with loose arrangement and great difference in size, and the coefficient of variation was high. While in the Yangdao 6, most of the amyloplasts were single, well developed, polyhedral, crystalline and compactly arranged, and the coefficient of variation was low. The amyloplasts in the dorsal region of endosperm were developed better than those in the ventral and central regions. The chalkiness in the endosperm resulted from badly-developed and loose-arranged starch granules, which was closely relevant to the transport of filling materials.