Identification of Green-Revertible Yellow 3(GRY3),encoding a 4-hydroxy-3-methylbut-2-enyl diphosphate reductase involved in chlorophyll synthesis under high temperature and high light in rice

Chlorophyll,a green pigment in photosynthetic organisms,is generated by two distinct biochemical pathways,the tetrapyrrole biosynthetic pathway(TBP) and the methylerythritol 4-phosphate(MEP)pathway.MEP is one of the pathways for isoprenoid synthesis in plants,with 4-hydroxy-3-methylbut-2-enyl diphosphate reductase(HDR) catalyzing its last step.In this study,we isolated a greenrevertible yellow leaf mutant gry3 in rice and cloned the GRY3 gene,which encodes a HDR participating in geranylgeranyl diphosphate(GGPP) biosynthesis in chloroplast.A complementation experiment confirmed that a missense mutation(C to T) in the fourth exon of LOC_Os03g52170 causes the gry3 phenotype.Under high temperature and high light,transcript and protein abundances of GRY3 were reduced in the gry3 mutant.Transcriptional expression of chlorophyll biosynthesis,chloroplast development,and genes involved in photosynthesis were also affected.Excessive reactive oxygen species accumulation,cell death,and photosynthetic proteins degradation were occurred in the mutant.The content of GGPP was reduced in gry3 compared with Nipponbare,resulting in a stoichiometric imbalance of tetrapyrrolic chlorophyll precursors.These results shed light on the response of chloroplast biogenesis and maintenance in plants to high-temperature and high-light stress.

Development of Chinese Near-isogenic Line of Rice and Their Differentiating Ability of Pathogenic Races of Blast Fungus

The rice( Oryza L. ) variety, Lijiangxintuanheigu(LTH), is a universally susceptible variety. A set of near-isogenic lines (NILs) of rice which has single resistance gene for each line was developed by use of LTH as recurrent parent and Kusabue,Tsuyuake, K1, PiNo. 4, K60 and BL1, which are from Kiyosawa's differential varieties(KDV), as donor parents. Differentiating ability of these NILs to Philippine isolates was compared with that of their donor parents, KDV and IRRI' s NILs. The results of comparative studies were as follows: (1) Differentiating ability of Chinese NILs to Philippine isolates was much stronger than that of their donor parents and that of KDV. (2) Chinese NILs had similar ability to differentiate races to IRRI's NILs or higher differentiating ability than them. But IRRI's NILs almost had no differentiating ability to isolates from japonica rice-growing regions. They can not be used to differentiate races of isolates from japonica ricegrowing regions. Whereas, Chinese NILs can be used to differentiate races of isolates from both japonica ricegrowing regions and indica rice-growing regions. It was concluded that the set of Chinese NILs can be used as international differentials.

Mapping and genetic validation of a grain size QTL qGS7.1 in rice(Oryza sativa L.)

Grain size is a major determinant of grain weight, which is one of the components of rice yield. The objective o this study was to identify novel, and important quantitative trait loci(QTLs) for grain size and weight in rice. QTLs were mapped using a BC4F4 population including 192 backcross inbred lines(BILs) derived from a backcross between Xiaolijing(XLJ) and recombinant inbred lines(RILs). The mapping population was planted in both Lingshui(Hainan, 2015) and Fuyang(Zhejiang, 2016), with the short-and long-day conditions, respectively. A total of 10 QTLs for grain length, four for grain width, four for the ratio of grain length to width, and 11 for grain weight were detected in at least one environment and were distributed across 11 chromosomes. The phenotypic variance explained ranged from 6.76–25.68%, 14.30–34.03%, 5.28–26.50%, and 3.01–22.87% for grain length, grain width, the ratio of grain length to width, and thousand grain weight, respectively. Using the sequential residual heterozygotes(SeqRHs) method, qGS7.1, a QTL for grain size and weight, was mapped in a 3.2-Mb interval on chromosome 7. No QTLs about grain size and weight were reported in previous studies in this region, providing a good candidate for functional analysis and breeding utilization.

Developmental Status Quo and Trends of Low-carbon Agriculture

In order to reduce carbon emission in agricultural production,this paper has discussed the developmental trends of low-carbon agriculture in terms of developing precision agriculture,improving the efficiency of fertilizer utilization,scientific use of pesticides,water-saving irrigation,ecological control of pests and diseases,as well as energy conservation and emission reduction by agricultural machinery and other agricultural practices.

A New PCR/LDR-Based Multiplex Functional Molecular Marker for Marker-Assisted Breeding in Rice

Marker-assisted selection or marker-aided selection(MAS)provides an effective complementary approach for conventional rice breeding with precise and speedy mobilization of target genes into elite genetic backgrounds.The targeted genes,however,may not be selected in the course of MAS due to the occasional recombination between the marker and the target gene/QTL during the many cycles of meiosis involved in breeding programs.This leads to failure in the selection of target traits(Gopalakrishnan et al,2008).

OstMAPKKK5, a truncated mitogen-activated protein kinase kinase kinase 5, positively regulates plant height and yield in rice

Rice(Oryza sativa L.)is a major food crop worldwide.Plant height and yield are important agronomic traits of rice.Several genes regulating plant height and/or yield have been cloned.However,the molecular mechanisms coordinating plant height and yield are not fully characterized.Here,we report a novel gene,OstMAPKKK5 that encodes a truncated variant of a mitogen-activated protein kinase kinase kinase 5(OsMAPKKK5)lacking an intact kinase domain.Transgenic plants overexpressing OstMAPKKK5 in indica cultivar 9311 showed increased plant height,grain length,grain width,1000-grain weight,grain number per main panicle,and yield.Real-time quantitative PCR showed that OstMAPKKK5 was widely expressed in various tissues and developmental stages.The increased plant height and yield were attributed to enlarged cell size.Overexpression of OstMAPKKK5 led to higher contents of various forms of endogenous gibberellin(GA),especially the most common active forms,GA1,GA3,GA4.We concluded that OstMAPKKK5 positively regulates plant height and yield in rice by affecting cell size,and that its underlying mechanism is based on increased endogenous GA content.

Genetic Variation of Inbred Lines of Maize Detected by SSR Markers

Simple sequence repeats (SSRs) were used to detect genetic variation among 21 maize(Zea mays L. ) inbred lines. Forty-three SSR primers selected from 69 primers gave stable amplification profiles, which could be clearly resolved on 3% Metaphor agarose gel, and produced 127 polymorphic amplified fragments.The average number of alleles per SSR locus was 2.95 with a range from 2 to 7. The polymorphism information content (PIC) for the SSR loci varied from 0.172 to 0.753 with an average of 0.511. Genetic similarities among the 21 lines ranged from 0.480 between the combination of Zhongzi451 vs. K12 up to 0.768 between CA156 vs. Ye478. The cluster analysis showed that 21 inbred lines could be classified into two distinct clusters with several subclusters, which corresponded to the heterotic groups determined by their pedigree information.Eight SSR primers, which had high level of polymorphism, could allow a rapid and efficient identification of 21 inbreds. Consequently, SSR markers could be used for measuring genetic variation of maize inbred lines and assigning them to heterotic groups.

Polymorphism of Kunitz Trypsin Inhibitor Protein in Natural Populations of Wild Soybean in Hebei

Hebei Province is one of the main distribution areas growing wild soybean( Glycine soja ) in China. In this study, 461 seed samples,collected from 18 natural populations in this province, were used to electrophoretically observe the change in forms and their frequencies of the Kunitz trypsin inhibitor protein (KTI)in individual populations and geographical areas. Allelic frequencies accounted for 85% for Tia and 15% for Tib in the total samples. Twelve populations examined were polymorphic at the KTI locus, accounting for over 50% in the populations investigated. Four populations, 22% of all the populations, were found to have natural cross-pollination with varied heterozygote rates of 3% -5.5%, and the average was 1% in the total sampies. Geographically, the mean Tib frequency in the north areas was higher than in the south, and higher in the mountainous area than in the plain areas. The populations in a lake ecological environment (Baiyangdian Lake) were almost monomorphic. No obvious relationship between the frequency and the geographical distance was observed. In addition, we first found a mutation for the absence of the KTI in wild soybean.

BREEDING OF DWARFING-STERILE WHEAT AND ITS POTENTIAL VALUES IN WHEAT BREEDING

A male sterile form of common wheat(Triticum aestivum) was found in China and it has been shown that the sterility is controlled by a single, dominant gene. The dominant Ms2 gene for male sterility (originally Tal assigned by Deng and Gao, 1982)is located on the short ann of chromosome 4D. This Ms2 gene has been extensively used as a

Integration and inheritance stability of foreign Bt toxin gene in the bivalent insect-resistant transgenic cotton plants

Genetic and expressional stability of Bt toxin gene is crucial for the breeding of insect-resistant transgenic cotton varieties and their commercialization. Genomic Southern blot analysis of R3, R4 and R5 generations of bivalent transgenic insect-resistant cotton plants was done in order to determine the integration, the copy number and the inheritance stability of Bt toxin gene in the transgenic cotton plants. The results indicated that there was a 4.7 kb positive band in the Southern blot when the genomic DNA of the bivalent transgenic insect-resistant cotton plants and the positive control (the plasmid) were digested with HindⅢ respectively. This result proved that the Bt toxin gene had been integrated into the genome of the cotton in full length. There is only one XhoⅠ restriction site in the Bt toxin gene. Southern blot analysis indicated that many copies of Bt toxin gene had been integrated into the genome of the cotton when the genomic DNA of transgenic plants was digested with XhoⅠ. Among them, there were four copies (about 17.7, 8, 5.5 and 4.7 kb in size) existing in all the tested plants of R3, R4 and R5 generations. The preliminary conclusion was that there were more than four copies of Bt toxin gene integrated into the genome of the cotton, among them, more than one copy can express and inherit steadily. This result provides a scientific basis for the breeding of the bivalent insect-resis- tant transgenic cotton plants and its commercialization.

TRANSFER OF ATRAZINE-RESISTANT GENE FROM NIGHTSHADE TO SOYBEAN CHLOROPLAST GENOME AND ITS EXPRESSION IN TRANSGENIC PLANTS

By binding the 32KD protein of chloroplast thylakoid membrane, the triazine herbicides such as atrazine inhibit the electron transport from the primary stable electron acceptor to thesecondary electron acceptor of photosystem Ⅱ and disturb the process of photosynthesis.A number of researches about the psbA gene coding the 32KD protein have been reported, but until now transfer of the atrazine-resistant psbA gene into the plant chloroplast genome has not yet succeeded. We cloned a chloroplast DNA fragment containing the psbA gene from an atrazine-resistant biotype of nightshade (Solanum nigrum) and analyzed

A STUDY ON CALLUS INDUCTION AND PLANTLET REGENERATION OF THE LEAF BLADE IN RICE (ORYZA SATIVA L. SUBSP. KENG)

The leaves of plant are a suitable material of protoplast isolation and culture. Hence, plantlet regeneration of the leaf tissue cell is the presupposition for protoplast culture and fusion. In recent year, the investigations of plantlets regenerated from leaf tissues in cultures have been greatly progressing in cereals, but no report on the induction and plantlet regeneration of calli has been seen in rice. This note deals with the results of this aspect.

Tagging Salt Tolerant Gene Using PCR Markers in Soybean

The purpose of this study was to screen and identify PCR markers associated with salt tolerant gene in soybean( Glycine soja L. ) so that salt tolerance can be identified efficiently and accurately. Between these tolerant and sensitivity to salt and three crosses were tested in this experiment. By BSA method, two codominant PCR markers were identified through the salt tolerant (sensitive) cuitivars bulks and the salt tolerant (sensitive) individual bulks of a F2 population. There was a 600bp band in the sensitive individuals and a 700bp band or two 700bp/600bp bands in the tolerant individuals. The markers were closely linked with salt tolerant/sensitive alleles. Moreover the markers were tested in the other two F2 populations from "salt tolerant cultivar × sensitive cuitivar" and confirmed by 12 salt tolerance cultivars and 13 salt sensitive cultivars with different genetic background. It indicated that the markers (700bp and 600bp) could be applied in salt tolerant identification of the soybean germplasm resources, and markers-assisted selection in salt tolerant breeding of soybean. The markers, its obtained method and application were patented for invention in 1998.

DEVELOPMENT OF COMMON WHEAT GERM-PLASM RESISTANT TO BARLEY YELLOW DWARF VIRUS BY BIOTECHNOLOGY

Barley yellow dwarf virus (BYDV) is one of the most serious wheat diseases in China. So far no resistance has been described in common wheat. A certain level of BYDV resistance was found in thirteen Triticeae species. Thinopyrum intermedium, two octoploids derived from TH. intermedium/wheat, Zhong 4 awnless and TAF46, and one disomic addition line, L1 derived from TAF46, showed good resistance to BYDV by enzyme linked immunosorbent assay (ELISA). Two wheat/TA. intermedium translocation lines, CPI 119880 and CPI 119899, showing good BYDV resistance were developed from L1 by using both CSph mutant and tissue culture. It is found that their BYDV resistance was controlled by a single dominant gene. Two cDNA probes pEleAcc3 and pPJN8 (E1-T1) were screened for detecting Th. intermedium DNA in wheat background. A specific band for the DNA of Th. intermedium and its derivatives was found in Southern hybridization. It is also possible to determine the size of the alien segment by comparing the relative density

Introduction of a rice blight resistance gene, Xa21, into five Chinese rice varieties through an Agrobacterium -mediated system

A cloned gene, Xa21 was transferred into five widely-used Chinese rice varieties through an Agrobacterium-mediated system, and over 110 independent transgenic lines were obtained. PCR and Southern analysis of transgenic plants revealed the integration of the whole Xa21 gene into the host genomes. The integrated Xa21 gene was stably inherited, and segregated in a 3 : 1 ratio in the selfed T1 generation when one copy of the gene was integrated in the transfor-mants. Inoculation tests displayed that transgenic T0 plants and Xa21 PCR-positive T1 plants were highly resistant to bacterial blight disease. The selected Xa21 homozygous resistant transgenic lines with desirable qualities may be propagated as new varieties or utilized in hybrid rice breeding.

Nucleolus organizer regions (Nor loci) of Chinese wheats

Nucleolus organizer regions (Nor loci) of a range of Chinese wheat landraces and cultivars (Triticum aestivum L. em Thell.) were analysed using genomic DNA extracted from leaves. Only two allelic variants of the Nor-B1 locus were found on chromosome 1B (Nor-B1a and Nor-B1g), while Nor-B1g was probably introduced from North America in the early 1960s. The even more recent introduction of the rye allele Nor-R1 in the early 1980s was also revealed. Eight allelic variants of the Nor-B2 locus on chromosome 6B (Nor-B2a, b, d, f, h, o, p and s) were identified. A Chinese origin for the a, d, f, o, p and s alkies is evident although the d allele was successfully introduced into Australian wheats in the early 1900s. Nor-B2h and Nor-B2b are again very recent introductions into Chinese wheat breeding programs, the former from CIMMYT wheats and the latter in association with the introduction of the 1RS/1BL translocation from Europe. On the basis of the presence of different combinations of Nor-B1 and Nor-B2

TRANSGENIC RICE PLANT OF A SUPERIOR CHINESE CULTIVAR Zhonghua NO. 11 CONTAINING THE B. t. δ-ENDOTOXIN GENE IN ITS GENOME

The B. t. δ-endotoxin protein gene has been transferred into a commercial rice variety Zhonghua No. 11 by the pollen-tube pathway method. The expression of β-glucuronidase activity in the transgenic plant （D3） was confirmed by histochemical analysis. Since the GUS gene was translationally fused with the B. t. δ-endotoxin protein gene in the recombinant plasmid pGYCK63, the expression of the GUS gene offered evidence for the expression of the δ-endotoxin protein gene in the transgenic rice plant.

Auxin signaling module OsSK41-OsIAA10-OsARF regulates grain yield traits in rice

Auxin is an important phytohormone in plants, and auxin signaling pathways in rice play key roles in regulating its growth, development, and productivity. To investigate how rice grain yield traits are regulated by auxin signaling pathways and to facilitate their application in rice improvement, we validated the functional relationships among regulatory genes such as OsIAA10, OsSK41, and OsARF21 that are involved in one of the auxin(OsIAA10) signaling pathways. We assessed the phenotypic effects of these genes on several grain yield traits across two environments using knockout and/or overexpression transgenic lines.Based on the results, we constructed a model that showed how grain yield traits were regulated by OsIAA10 and OsTIR1, OsAFB2, and OsSK41 and OsmiR393 in the OsSK41-OsIAA10-OsARF module and by OsARF21 in the transcriptional regulation of downstream auxin response genes in the OsSK41-OsIAA10-OsARF module. The population genomic analyses revealed rich genetic diversity and the presence of major functional alleles at most of these loci in rice populations. The strong differentiation of many major alleles between Xian/indica and Geng/japonica subspecies and/or among modern varieties and landraces suggested that they contributed to improved productivity during evolution and breeding. We identified several important aspects associated with the genetic and molecular bases of rice grain and yield traits that were regulated by auxin signaling pathways.We also suggested rice auxin response factor(OsARF) activators as candidate target genes for improving specific target traits by overexpression and/or editing subspecies-specific alleles and by searching and pyramiding the ‘best' gene allelic combinations at multiple regulatory genes in auxin signaling pathways in rice breeding programs.