2021, Volume 9, Issue 4

20 August 2021
  
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  • Reviews
    Nigarin Sultana, Shahidul Islam, Angela Juhasz, Wujun Ma
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    Wheat leaf senescence is a developmental process that involves expressional changes in thousands of genes that ultimately impact grain protein content (GPC), grain yield (GY), and nitrogen use efficiency. The onset and rate of senescence are strongly influenced by plant hormones and environmental factors e.g. nitrogen availability. At maturity, decrease in nitrogen uptake could enhance N remobilization from leaves and stem to grain, eventually leading to leaf senescence. Early senescence is related to high GPC and somewhat low yield whereas late senescence is often related to high yield and somewhat low GPC. Early or late senescence is principally regulated by up and down-regulation of senescence associated genes. Integration of external and internal factors together with genotypic variation influence senescence associated genes in a developmental age dependent manner. Although regulation of genes involved in senescence has been studied in rice, Arabidopsis, maize, and currently in wheat, there are genotype-specific variations yet to explore. A major effort is needed to understand the interaction of positive and negative senescence regulators in determining the onset of senescence. In wheat, increasing attention has been paid to understand the role of positive senescence regulator, e.g. GPC-1, regulated gene network during early senescence time course. Recently, gene regulatory network involved early to late senescence time course revealed important senescence regulators. However, the known negative senescence regulator TaNAC-S gene has not been extensively studied in wheat and little is known about its value in breeding. Existing data on senescence-related transcriptome studies and gene regulatory network could effectively be used for functional study in developing nitrogen efficient wheat varieties.

  • Reviews
    Xiaofei Xue, Shangyi Du, Fuchao Jiao, Menghan Xi, Aiguo Wang, Haicheng Xu, Qiqing Jiao, Xin Zhang, Hao Jiang, Jingtang Chen, Ming Wang
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    Seed germination is the process by which an organism grows from a seed. It requires suitable conditions and environmental factors. Maize is one of the most important crops worldwide. Germination influences both final maize yield and quality. Seed germination is regulated by a complex gene network. It is also influenced by endogenous (phytohormones and nutrients) and exogenous (temperature and water) inputs, involving molecular networks only partly identified to date. This review describes current understanding of the influence of temperature, water, phytohormones, and nutrients in regulating maize seed germination, and indicates knowledge gaps that should be addressed.

  • Research Papers
    Shijuan Yan, Qing Liu, Thomas Naake, Wenjie Huang, Mengyu Chen, Qian Kong, Sheng Zhang, Wenyan Li, Xuan Li, Qinjian Liu, Jianyuan Yang, Alisdair R. Fernie, Bin Liu
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    Rice with panicle-blast resistance is needed for stable rice production. Although we have previously demonstrated that OsGF14b underlies a quantitative trait locus that positively regulates rice panicle blast resistance, the mechanism is still unknown. In this study, a multi-omics approach was used to investigate the possible downstream signaling pathway regulated by OsGF14b. OsGF14b both strongly activated the gibberellin biosynthetic pathway during pathogen infection and reprogrammed the lignin biosynthetic pathway. Reduced lignin accumulation was observed in glumes of OsGF14b-overexpressing plants in comparison with the wild type after pathogen inoculation. OsGF14b activated the auxin and jasmonic acid signaling pathways, but inactivated the salicylic acid signaling pathway. Auxin and jasmonic acid appeared to act independently on OsGF14b-mediated panicle blast resistance. The roles of gibberellin, lignin, and auxin were different from their roles in leaf blast, suggesting that different mechanisms underlie leaf and panicle blast resistance in rice. This study provides a comprehensive catalog of molecular changes that could be targets for future studies of rice panicle blast resistance.

  • Research Papers
    Tong Zheng, Chen Hua, Lei Li, Zhengxi Sun, Minmin Yuan, Guihua Bai, Gavin Humphreys, Tao Li
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    Fusarium head blight (FHB) is a global wheat disease that devastates wheat production. Resistance to FHB spread within a wheat spike (type II resistance) and to mycotoxin accumulation in infected kernel (type III resistance) are the two main types of resistance. Of hundreds of QTL that have been reported, only a few can be used in wheat breeding because most show minor and/or inconsistent effects in different genetic backgrounds. We describe a new strategy for identifying robust and reliable meta-QTL (mQTL) that can be used for improvement of wheat FHB resistance. It involves integration of mQTL analysis with mQTL physical mapping and identification of single-copy markers and candidate genes. Using meta-analysis, we consolidated 625 original QTL from 113 publications into 118 genetic map-based mQTL (gmQTL). These gmQTL were further located on the Chinese Spring reference sequence map. Finally, 77 high-confidence mQTL (hcmQTL) were selected from the reference sequence-based mQTL (smQTL). Locus-specific single nucleotide polymorphism (SNP) and simple sequence repeat (SSR) markers and 17 genes responsive to FHB were then identified in the hcmQTL intervals by combined analysis of transcriptomic and proteomic data. This work may lead to a comprehensive molecular breeding platform for improving wheat resistance to FHB.

  • Research Papers
    Lei Chen, Gang Gu, Chengxu Wang, Zhufeng Chen, Wei Yan, Man Jin, Gang Xie, Junli Zhou, Xing Wang Deng, Xiaoyan Tang
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    Herbicide resistance in crop plants is valuable for integrated weed management in agriculture. Herbicide resistant rice, in particular, is important to management of weedy rice, a close relative of cultivated rice and a noxious weed prevalent in rice fields that remains challenging to farmers worldwide. Herbicide resistant plants can be obtained through transgenic approach or by mutagenesis of regular plant and screening of mutants with elevated resistance to herbicide. In this study, we conducted ethyl methyl sulfonate mutagenesis (EMS) to elite indica cultivar Huanghuazhan (HHZ) and screened for mutants resistant to imazapic, a herbicide that can inhibit the acetolactate synthase (ALS) in plants. We obtained three mutants of OsALS gene that have not been reported previously in rice. One of the mutants, with Trp548 changed to Met (W548M), was analyzed in more details in this study. This mutation had no negative effect on the plant physiology and morphology as well as rice yield. Compared with the imidazolinone-resistant mutant S627N (Ser627 changed to Asn) that has been deployed for Clearfield rice development, W548M mutant showed high levels of resistance to a broad spectrum of five families of ALS-inhibiting herbicides, in addition to a higher level of resistance to herbicides of the imidazolinone family. The herbicide-resistance was stably inherited by crossing into other rice lines. Thus, the W548M mutation provides a valuable resource for breeding of herbicide resistant rice and weed management.

  • Research Papers
    Xinmin Hu, Guihua Wang, Xuemei Du, Hongwei Zhang, Zhenxiang Xu, Jie Wang, Guo Chen, Bo Wang, Xuhui Li, Xunji Chen, Junjie Fu, Jun Zheng, Jianhua Wang, Riliang Gu, Guoying Wang
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    Drought is one of the most critical abiotic stresses influencing maize yield. Improving maize cultivars with drought tolerance using marker-assisted selection requires a better understanding of its genetic basis. In this study, a doubled haploid (DH) population consisting of 217 lines was created by crossing the inbred lines Han 21 (drought-tolerant) and Ye 478 (drought-sensitive). The population was genotyped with a 6 K SNP assay and 756 SNP (single nucleotide polymorphism) markers were used to construct a linkage map with a length of 1344 cM. Grain yield (GY), ear setting percentage (ESP), and anthesis-silking interval (ASI) were recorded in seven environments under well-watered (WW) and water-stressed (WS) regimes. High phenotypic variation was observed for all traits under both water regimes. Using the LSMEAN (least-squares mean) values from all environments for each trait, 18 QTL were detected, with 9 associated with the WW and 9 with the WS regime. Four chromosome regions, Chr. 3: 219.8-223.7 Mb, Chr. 5: 191.5-194.7 Mb, Chr. 7: 132.2-135.6 Mb, and Chr. 10: 88.2-89.4 Mb, harbored at least 2 QTL in each region, and QTL co-located in a region inherited favorable alleles from the same parent. A set of 64 drought-tolerant BC3F6 lines showed preferential accumulation of the favorable alleles in these four regions, supporting an association between the four regions and maize drought tolerance. QTL-by-environment interaction analysis revealed 28 edQTL (environment-dependent QTL) associated with the WS regime and 22 associated with the WW regime for GY, ESP, and ASI. All WS QTL and 55.6% of WW QTL were located in the edQTL regions. The hotspot genomic regions identified in this work will support further fine mapping and marker-assisted breeding of drought-tolerant maize.

  • Research Papers
    Zhaobo Li, Qun Cheng, Zhuoran Gan, Zhihong Hou, Yuhang Zhang, Yongli Li, Haiyang Li, Haiyang Nan, Cen Yang, Linnan Chen, Sijia Lu, Wenqian Shi, Liyu Chen, Yanping Wang, Chao Fang, Liping Kong, Tong Su, Shichen Li, Kun Kou, Lingshuang Wang, Fanjiang Kong, Baohui Liu, Lidong Dong
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    Flowering time is an important agronomic trait for soybean yield and adaptation. However, the genetic basis of soybean adaptation to diverse latitudes is still not clear. Four NIGHT LIGHT-INDUCIBLE AND CLOCK-REGULATED 2 (LNK2) homeologs of Arabidopsis thaliana LNK2 were identified in soybean. Three single-guide RNAs were designed for editing the four LNK2 genes. A transgene-free homozygous quadruple mutant of the LNK2 genes was developed using the CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR-associated protein 9). Under long-day (LD) conditions, the quadruple mutant flowered significantly earlier than the wild-type (WT). Quantitative real-time PCR (qRT-PCR) revealed that transcript levels of LNK2 were significantly lower in the quadruple mutant than in the WT under LD conditions. LNK2 promoted the expression of the legume-specific E1 gene and repressed the expression of FT2a. Genetic markers were developed to identify LNK2 mutants for soybean breeding. These results indicate that CRISPR/Cas9-mediated targeted mutagenesis of four LNK2 genes shortens flowering time in soybean. Our findings identify novel components in flowering-time control in soybean and may be beneficial for further soybean breeding in high-latitude environments.

  • Research Papers
    Dexin Liu, Xueying Liu, Yao Su, Xiao Zhang, Kai Guo, Zhonghua Teng, Jian Zhang, Dajun Liu, Zhengsheng Zhang
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    Naturally colored cotton fiber is environment-friendly but has monotonous color and poor fiber quality. Identification of green fiber or fuzz genes would aid in investigating the biosynthesis of green pigments in cotton fibers. In this study, we established a mapping population and found that the Lgf trait (white lint and green fuzz) from Gossypium hirsutum race latifolium is controlled by an incompletely dominant gene. The Lgf locus was mapped to a 71-kb interval on chromosome 21 containing seven genes, including a transcription factor with similarity to Arabidopsis MYB9. Harboring 13 SNPs and a 4-bp insertion/deletion in its promoter, GhMYB9 was highly up-regulated in the critical period for green pigment development in fuzz. Virus-induced gene silencing of GhMYB9 in a green-fuzz accession of G. hirsutum race latifolium TX-41 conferred white or light green fuzz. These results suggest that GhMYB9 is an important contributor to green pigments in cotton fiber and shed light on the regulatory mechanism controlling green pigmentation.

  • Research Papers
    Jun Chen, Yan Gong, Yuan Gao, Yongbin Zhou, Ming Chen, Zhaoshi Xu, Changhong Guo, Youzhi Ma
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    NAC family transcription factors (TFs) are important regulators in plant development and stress responses. However, the biological functions of NAC TFs in wheat are rarely studied. In this study, 43 putative drought-induced NAC genes were identified from de novo transcriptome sequencing data of wheat following drought treatment. Twelve wheat NACs along with ten known stress-related NACs from Arabidopsis and rice were clustered into Group II based on a phylogenetic analysis. TaNAC48, which showed a higher and constitutive expression level in Group II, was selected for further investigation. TaNAC48 transcript was up-regulated by drought, PEG, H2O2 and abscisic acid (ABA) treatment and encoded a nuclear localized protein. Overexpression of TaNAC48 significantly promoted drought tolerance with increased proline content, and decreased rates of water loss, malondialdehyde (MDA), H2O2 and O2 content. Root length and a stomatal aperture assay confirmed that TaNAC48-overexpression plants increased sensitivity to ABA. Electrophoretic mobility shift assay (EMSA) and luciferase reporter analysis indicated that TaAREB3 could bind to a cis-acting ABA-responsive element (ABRE) on TaNAC48 promoter and activate the expression of TaNAC48. These results suggest that TaNAC48 is essential in mediating crosstalk between the ABA signaling pathway and drought stress responses in wheat.

  • Research Papers
    Xichao Sun, Wei Ren, Peng Wang, Fanjun Chen, Lixing Yuan, Qingchun Pan, Guohua Mi
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    Nitrogen (N) deficiency is one of the main factors limiting maize (Zea mays L.) productivity. Genetic improvement of root traits could improve nitrogen use efficiency. An association panel of 461 maize inbred lines was assayed for root growth at seedling emergence under high-nitrate (HN, 5 mmol L−1) and low-nitrate (LN, 0.05 mmol L−1) conditions. Twenty-one root traits and three shoot traits were measured. Under LN conditions, the root-to-shoot ratio, root dry weight, total root length, axial root length, and lateral root length on the primary root were all increased. Under LN conditions, the heritability of the plant traits ranged from 0.43 to 0.82, a range much wider than that of 0.27 to 0.55 observed under HN conditions. The panel was genotyped with 542,796 high-density single-nucleotide polymorphism (SNP) markers. Totally 328 significant SNP markers were identified using either mixed linear model (MLM) or general linear model analysis, with 34 detected by both methods. In the 100-kb intervals flanking these SNP markers, four candidate genes were identified. Under LN conditions, the protoporphyrinogen IX oxidase 2 gene was associated with total root surface area and the DELLA protein-encoding gene was associated with the length of the visible lateral root zone of the primary root. Under HN conditions, a histone deacetylase gene was associated with plant height. Under both LN and HN conditions, the gene encoding MA3 domain-containing protein was associated with the first whorl crown root number. The phenotypic and genetic information from this study may be exploited for genetic improvement of root traits aimed at increasing NUE in maize.

  • Research Papers
    Jin Chen, Xiangcheng Zhu, Jiang Xie, Guoqiang Deng, Tianhua Tu, Xianjiao Guan, Zhen Yang, Shan Huang, Xianmao Chen, Caifei Qiu, Yinfei Qian, Caihong Shao, Minggang Xu, Chunrui Peng
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    Rational nitrogen (N) application can greatly increase rice (Oryza sativa L.) yield. However, excessive N input can lead not only to low N use efficiency (NUE) but also to severe environmental pollution. Reducing N application rate with a higher planting density (RNHD) is recommended to maintain rice yield and improve NUE. The effects of RNHD on fertilizer N fate and rice root growth traits remain unclear. We accordingly conducted a two-year field experiment to investigate the influence of RNHD on rice yield, fertilizer 15N fate, and root growth in a double-rice cropping system in China. In comparison with the conventional practice of high N application with sparse planting, RNHD resulted in similar yield and biomass production as well as plant N uptake. RNHD increased agronomic NUEs by 23.3%-31.9% (P < 0.05) and N recovery efficiency by 17.4%-24.1% (P < 0.05). RNHD increased fertilizer 15N recovery rate by 14.5%-34.7% (P < 0.05), but reduced 15N retention rate by 9.2%-12.0% (P < 0.05). Although a reduced N rate led to significantly reduced root length, surface area, volume, and biomass, these root traits were significantly increased by higher planting density. RNHD did not affect these root morphological traits and reduced activities of nitrate reductase (NR) and glutamine synthetase (GS) only at tillering stage. Plant N uptake was significantly positively correlated with these root traits, but not correlated with NR and GS activities. Together, these findings show that reducing N application with dense planting can lead to high plant N uptake by maintaining rice root growth and thus increase NUE.

  • Research Papers
    Xia Zhang, Caihong Cui, Yinguang Bao, Honggang Wang, Xingfeng Li
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    Thinopyrum intermedium has been used as a resource for improving resistance to biotic and abiotic stresses and yield potential in common wheat. Wheat line SN304 was derived from a cross between common wheat cultivar Yannong 15 and Th. intermedium. Genomic in situ hybridization (GISH) produced no hybridization signal in SN304 using Th. intermedium genomic DNA as a probe, but fluorescence in situ hybridization (FISH) using oligonucleotides AFA-3, AFA-4, pAs1-1, pAs1-3, pAs1-4, pAs1-6, pSc119.2-1, and (GAA)10 as probes detected hybridization signals on chromosomes 2A, 7A, 2B, 3B, 6B, and 7B in SN304 that differed from Yannong 15. Results of specific markers also indicated that there were Th. intermedium chromatin introgressions on different chromosomes in SN304. In a hydroponic culture experiment, SN304 not only produced more biomass and higher stem and leaf dry weight but also accumulated more phosphorus than Yannong 15 under phosphorus-deficiency stress. Moreover, SN304 produced a lower pH and released more organic acids, especially oxalic acid, than Yannong 15, which suggests that SN304 exudates enabled more absorbance of P than Yannong 15 under comparable conditions. The results indicate that SN304 is a wheat-Th. intermedium introgression line with tolerance to phosphorus-deficiency stress.

  • Research Papers
    Bin Chen, Yan Zhang, Jun Yang, Man Zhang, Qingming Ma, Xingfen Wang, Zhiying Ma
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    Verticillium wilt, a devastating disease in cotton caused by Verticillium dahliae, reduces cotton quality and yield. Heterotrimeric GTP-binding proteins, consisting of Gα, Gβ, and Gγ subunits, transducers of receptor signaling, function in a wide range of biological events. However, the function of Gα proteins in the regulation of defense responses in plants is largely unexplored, except for a few reports on model species. In the present study, a cotton G-protein α-subunit-encoding gene (GhGPA) was isolated from Verticillium wilt-resistant Gossypium hirsutum (upland cotton) cv. ND601. GhGPA transcription was up-regulated under V. dahliae stress, with higher expression in tolerant than in susceptible cotton cultivars. Subcellular localization revealed GhGPA to be located in the plasma membrane. GhGPA shows high (85.0%) identity with Arabidopsis AT2G26300 (AtGPA1), and AtGPA1 gpa1-4 mutants displayed susceptibility to V. dahliae. Ectopic expression of GhGPA successfully restored the resistance of Arabidopsis gpa1-4 mutants to Verticillium wilt and made them more resistant than the wild type. Overexpression of GhGPA in Arabidopsis markedly increased the resistance and resulted in dramatic up-regulation of pathogenesis-related (PR) genes and increased in H2O2 accumulation and salicylic acid (SA) and jasmonic acid (JA) contents. However, suppressing GhGPA expression via virus-induced gene silencing (VIGS) increased susceptibility to Verticillium wilt, down-regulated the expression of PR and marker genes in SA and JA signaling pathways, and reduced H2O2 content. The contents of SA and JA in Arabidopsis gpa1-4 and VIGS cotton were lower than those in the wild type and empty-vector control. However, GhGPA-overexpressing Arabidopsis contained more SA and JA than the wild type when inoculated with V. dahliae. Thus, GhGPA plays a vital role in Verticillium wilt resistance by inducing SA and JA signaling pathways and regulating the production of reactive oxygen species. These findings not only broaden our knowledge about the biological role of GhGPA, but also shed light on the defense mechanisms involving GhGPA against V. dahliae in cotton.

  • Research Papers
    Jialing Lu, Chunchao Wang, Fan Zhang, Dan Zeng, Yongli Zhou
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    Bacterial blight (BB), which is caused by Xanthomonas oryzae pv. oryzae (Xoo), is one of the most destructive bacterial diseases of rice (Oryza sativa L.). During plant defense responses, microRNAs (miRNAs) play important roles in regulating disease resistance. However, the functions of miRNAs in the interaction between rice and Xoo remain relatively uncharacterized. In this study, we compared the miRNA profiles of the BB resistant rice introgression line F329 and its susceptible recurrent parent Huang-Hua-Zhan (HHZ) at multiple time points after inoculation with Xoo. A total of 538 known and 312 novel miRNAs were identified, among which only 17 and 26 were responsive to Xoo infection in F329 and HHZ, respectively. Compared with the expression levels in HHZ, 37 up-regulated and 53 down-regulated miRNAs were detected in F329. The predicted target genes for the miRNAs differentially expressed between F329 and HHZ were revealed to be associated with flavonoid synthesis, the reactive oxygen species regulatory pathway, plant hormone signal transduction, defense responses, and growth and development. Additionally, the patterns of interactions between osa-miR390-3p, novel_miR_104, novel_miR_238, osa-miR166k-5p, osa-miR529b, and osa-miR167h-3p and their target genes were further validated by quantitative real-time PCR. Furthermore, we overexpressed osa-miR167h-3p in transgenic plants and proved that this miRNA positively regulates the resistance of rice to BB. These results provide novel information regarding the miRNA-based molecular mechanisms underlying the disease resistance of rice. The data presented herein may be useful for engineering rice BB resistance via miRNAs.

  • Research Papers
    Na Cui, Xiaofeng Chen, Yan Shi, Meirong Chi, Jiantuan Hu, Kunlong Lai, Zonghua Wang, Haifeng Wang
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    DNA methylation participates in regulating the expression of coding and non-coding regions in plants. To investigate the association between DNA methylation and pathogen infection, we used whole-genome bisulfite sequencing to survey temporal DNA methylation changes in rice after infection with the rice blast fungus Magnaporthe oryzae. In contrast to previous findings in Arabidopsis, global DNA methylation levels in rice increased slightly after rice blast infection. We identified over 38,000 differentially methylated regions (DMRs), and hypermethylated DMRs far outnumbered hypomethylated DMRs. Most DMRs were located in transposable element regions. Using transcriptome analysis, we identified 8830 differentially expressed genes (DEGs) after 1, 3, and 5 days of infection. Over one-third of DEGs, most of which were CHH-type DMRs, were associated with DMRs. Functional analysis of the CHH DMR-DEGs indicated their involvement in many important biological processes, including cell communication and response to external stimulus. The transcription of many NBS-LRR family genes was affected by changes in DNA methylation, suggesting that DNA methylation plays essential roles in the response of rice to M. oryzae infection. More broadly, the DNA methylation analysis presented here sheds light on epigenomic involvement in plant defense against fungal pathogens.

  • Research Papers
    Hong Zhou, Wei Luo, Shang Gao, Jian Ma, Meixue Zhou, Yuming Wei, Youliang Zheng, Yaxi Liu, Chunji Liu
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    Kernel weight (KW), together with kernel number per unit area, determines yield of cereal crops. Here, two barley recombinant inbred lines (RILs) populations with a shared parent were used to identify loci controlling KW. One is Baudin/AWCS276 (BA) for which a linkage map was available. Several large-effect QTL controlling KW were detected in this population. Another is Morex/AWCS276 (MA). A linkage map with 5273 makers formed 1454 clusters, was constructed by the genotyping by sequence (GBS) data of 201 RILs from this population. A single marker was selected to represent each of the clusters and the linkage map constructed with these markers covers a total length of 1022.4 cM with an average interval of approximately 0.7 cM between loci. Three of the large-effect loci controlling KW (located on 2HL, 6HL, and 7HL, respectively) identified from the BA population were also detected in the MA population under different environments. The locus on 6HL was detected in each of the experiments conducted for both populations thus was selected for developing near isogenic lines (NILs). Apart from KW, the two isolines for each pair of the putative NILs obtained showed no significant difference for any of the morphological characteristics assessed. The average difference in KW between the isolines for all the NILs obtained was about 15% based on assessments under both glasshouse and field conditions. Taken advantage that high quality genome assemblies for both Morex and AWCS276 are available, we identified candidate genes underlying two of the three loci based on an orthologous analysis. The NILs developed and the candidate genes identified in this study should facilitate the cloning and functional analysis of genes regulating KW in barley.

  • Research Papers
    Ana M. Casas, Carlota R. Gazulla, Arantxa Monteagudo, Carlos P. Cantalapiedra, Marian Moralejo, M. Pilar Gracia, Francisco J. Ciudad, William T.B. Thomas, Molina-Cano José L., Scott Boden, Bruno Contreras-Moreira, Ernesto Igartua
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    Response to vernalization and photoperiod are the main determinants controlling the time to flowering in temperate cereals. While the individual genes that determine a plant’s response to these environmental signals are well characterized, the combinatorial effect on flowering time of allelic variants for multiple genes remains unresolved. This study investigated the genetic control of flowering-time in a biparental population of spring barley, derived from a wide cross between a late-flowering European and an early-flowering North-American cultivar. While the major flowering time genes are not segregating in the Beka × Logan cross, large variation in flowering was observed. We identified five QTL, with both parents found to contribute early alleles. The catalog of QTL discovered aligns with several candidate genes affecting flowering time in barley. The combination of particular alleles at HvCEN, HvELF3 and HvFT1 in Logan are responsible for the earliness of this cultivar. Interestingly, earliness for flowering could be further enhanced, with Beka found to contribute three early alleles, including a QTL co-locating with a HvFD-like gene, suggesting that there are diverse aspects of the flowering-time pathway that have been manipulated in these two cultivars. Epistatic interactions between flowering-time QTL or candidate genes were observed in field data and confirmed under controlled conditions. The results of this study link photoperiod-dependent flowering-time genes with earliness per se genes into a single model, thus providing a unique framework that can be used by geneticists and breeders to optimize flowering time in barley.

  • Research Papers
    Qiuhong Wu, Yongxing Chen, Jingzhong Xie, Lingli Dong, Zhenzhong Wang, Ping Lu, Rongge Wang, Chengguo Yuan, Yan Zhang, Zhiyong Liu
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    Reduced plant height is one of the most important traits related to lodging resistance and crop yield. The use of reduced height genes has been one of the main features in breeding modern high-yielding wheat varieties with less lodging. A spontaneous dwarf mutant DD399 was identified in a high yielding, gibberellic acid (GA)-insensitive, lodging-resistant variety Nongda 399 (ND399). Significant differences in upper internode lengths between mutant DD399 and wild type ND399 were caused by reduced cell elongation. The plant height of ND399 × DD399 F1 hybrids was intermediate between the parents, indicating incomplete dominance or a dose-response effect of a reduced height gene. Plant height showed continuous distribution in the F2 population, and segregation distortion was observed among the 2292 F2:3 progenies. The reduced height mutation was characterized by Illumina 90 K iSelect SNP genotyping and bulked segregant RNA-Seq (BSR-Seq) analysis of the segregating population. A concentrated cluster of polymorphic SNPs associated with the reduced height phenotype was detected in the distal region of chromosome arm 2BL. Co-segregation of reduced height phenotype with the clustered markers revealed a 36 Mb terminal deletion of chromosome 2BL in mutant DD399.

  • Research Papers
    Ruiqi Zhang, Chuanxi Xiong, Huanqing Mu, Ruonan Yao, Xiangru Meng, Lingna Kong, Liping Xing, Jizhong Wu, Yigao Feng, Aizhong Cao
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    Powdery mildew, caused by the biotrophic fungus Blumeria graminis f. sp. tritici (Bgt), is a global disease that poses a serious threat to wheat production. To explore additional resistance gene, a wheat-Dasypyrum villosum 1V#5 (1D) disomic substitution line NAU1813 (2n = 42) with high level of seedling resistance to powdery mildew was used to generate the recombination between chromosomes 1V#5 and 1D. Four introgression lines, including t1VS#5 ditelosomic addition line NAU1815, t1VL#5 ditelosomic addition line NAU1816, homozygous T1DL·1VS#5 translocation line NAU1817, and homozygous T1DS·1VL#5 translocation line NAU1818 were developed from the selfing progenies of 1V#5 and 1D double monosomic line that derived from F1 hybrids of NAU1813/NAU0686. All of them were characterized by fluorescence in situ hybridization, genomic in situ hybridization, 1V-specific markers analysis, and powdery mildew tests at different developmental stages. A new powdery mildew resistance gene named Pm67 was physically located in the terminal bin (FL 0.70-1.00) of 1VS#5. Lines with Pm67 exhibited seedling stage immunity and tissue-differentiated reactions at adult plant stage. The sheaths, stems, and spikes of the Pm67 line were still immune, but the leaves showed a low degree of susceptibility. Microscopic observation showed that most penetration attempts were stopped in association with papillae on the sheath, and colonies cannot form conidia on the susceptible leaf of Pm67 line at adult plant stage, suggesting that the defence layers of the Pm67 line is tissue-differentiated. Thus, the T1DL·1VS#5 translocation line NAU1817 provides a new germplasm in wheat breeding for improvement of powdery mildew resistance.

  • Research Papers
    Raheel OsmanYan Zhu, Weixing Cao, Zhifeng Ding, Meng Wang, Leilei Liu, Liang Tang, Bing Liu
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    Extreme high-temperature stress (HTS) associated with climate change poses potential threats to wheat grain yield and quality. Wheat grain protein concentration (GPC) is a determinant of wheat quality for human nutrition and is often neglected in attempts to assess climate change impacts on wheat production. Crop models are useful tools for quantification of temperature impacts on grain yield and quality. Current crop models either cannot simulate or can simulate only partially the effects of HTS on crop N dynamics and grain N accumulation. There is a paucity of observational data on crop N and grain quality collected under systematic HTS scenarios to develop algorithms for model improvement as well as evaluate crop models. Two-year phytotron experiments were conducted with two wheat cultivars under HTS at anthesis, grain filling, and both stages. HTS significantly reduced total aboveground N and increased the rate of grain N accumulation, while total aboveground N and the rate of grain N accumulation were more sensitive to HTS at anthesis than at grain filling. The observed relationships between total aboveground N, rate of grain N accumulation, and HTS were quantified and incorporated into the WheatGrow model. The new HTS routines improved simulation of the dynamics of total aboveground N, grain N accumulation, and GPC by the model. The improved model provided better estimates of total aboveground N, grain N accumulation, and GPC under HTS (the normalized root mean square error was reduced by 40%, 85%, and 80%, respectively) than the original WheatGrow model. The improvements in the model enhance its applicability to the assessment of climate change effects on wheat grain quality by reducing the uncertainties of simulating N dynamics and grain quality under HTS.

  • Research Papers
    Yongli Luo, Wenqian Li, Cui Huang, Junhao Yang, Min Jin, Jin Chen, Dangwei Pang, Yonglan Chang, Yong Li, Zhenlin Wang
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    Drought at the grain filling stage of wheat will cause premature leaf senescence, thus leading to considerable loss of wheat yield. Therefore, this paper aims to establish a cultivation technology for strong drought resistance, delayed senescence, and yield improvement based on the analysis of hormones homeostasis obtained by applying chemical control substances. Experiments were conducted with two genotypes of wheat. Four water irrigation treatments were applied to impose the water deficit, including well-watered control treatment (WW), mild water deficit (MiWD), moderate water deficit (MoWD), and severe water deficit (SWD). Exogenous abscisic acid (ABA) was sprayed on the plants at the anthesis stage of the wheat. As a result, exogenous ABA reduced initial senescence rate (r0), total duration of chlorophyll (Chltotal), rapid senescence phase (Chlloss), and the accumulated temperature at an inflection point (M) but improved the persistence phase (Chlper) of flag leaves under all of the four treatments. However, exogenous ABA produced inconsistent effects on photoassimilate relocation and grain weight under different treatments. It produced positive regulatory effects on grain weight under WW, MiWD, and MoWD treatments. On the one hand, spraying ABA during the persistence phase of flag leaves reduced the ratios of zeatin to gibberellin (Z/GA3), spermine to spermidine (Spm/Spd), and salicylic acid to ABA (SA/ABA), which prolonged active photosynthesis by stimulating high level of proline (Pro) and increased the activities of antioxidant enzymes, such as superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and ascorbate peroxidase (APX). Therefore, drought tolerance was enhanced, and more photosynthetic assimilates were accumulated. On the other hand, the rapid senescence phase and the transport rate of assimilates into grains were accelerated, resulting in higher grain weight, yield, and water use efficiency (WUE). However, under SWD treatment, exogenous ABA improved the ratio of SA/ABA, leading to low Pro content and low antioxidant enzyme activity of flag leaves in the rapid loss phase. Meanwhile, drought resistance declined and the transport duration of assimilates into grains was shortened, thus making photosynthetic assimilates redundant. Therefore, exogenous ABA can lead to the reduction in grain weight, yield, and WUE of wheat under SWD treatment.

  • Research Papers
    Tengfei Lyu, Jie Shen, Jun Ma, Peng Ma, Zhiyuan Yang, Zou Dai, Chuangang Zheng, Min Li
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    Machine transplanting and the application of slow-release nitrogen (N) fertilizer (SRNF) have played vital roles in the modernization of rice production. We aimed to determine the effects of potted-seedling transplanting—a new machine-transplanting method—and SRNF on hybrid rice yields. A 2-year split-plot experiment (2016-2017) was conducted in Meishan, Sichuan province, China, using two machine-transplanting methods (potted-seedling and blanket-seedling) and three N treatments. Total green leaf area, high-effective leaf area and its rate at heading, net photosynthetic rate of flag leaves 7 days after heading, glutamate synthase (GOGAT) and glutamine synthase (GS) activity after heading, dry matter production, and N accumulation at heading and maturity increased under the potted-seedling method or 70% SRNF as a base + 30% urea application at the panicle initiation stage (SBUP). Stem diameter and number of small and of all vascular bundles at the neck-panicle node in potted-seedling plants increased as a result of increasing numbers of effective panicles, secondary branches, and spikelets. In potted-seedling plants, treatment with SBUP increased the number of large and total vascular bundles at the panicle-neck internode and the number of differentiated and surviving secondary branches and spikelets and decreased the number of ineffective tillers and degenerated secondary branches and spikelets. We conclude that the potted-seedling machine transplanting method and SRNF combined with urea topdressing can strengthen the source-sink relationship in rice, resulting in higher yields.

  • Research Papers
    Yuanyuan Zhang, Yang Cao, Hongying Zheng, Wenqi Feng, Jingtao Qu, Fengling Fu, Wanchen Li, Haoqiang Yu
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    Improved chilling tolerance is important for maize production. Previous efforts in transgenics and marker-assisted breeding have not achieved practical results. In this study, the antifreeze protein (AnAFP) from the super-xerophyte Ammopiptanthus nanus was aligned to KnS-type dehydrins. Phosphorylation in vitro and subcellular localization showed that AnAFP was phosphorylated by maize casein kinase II and translocated from nucleus to cytoplasm under chilling stress. AnAFP also increased lactate dehydrogenase activity. A parent line of commercial maize hybrids was transformed with the AnAFP gene. Based on thermal asymmetric interlaced PCR, one hemizygous and two homozygous integration sites were identified in one T1 line. Ectopic expression of AnAFP in transgenic lines was confirmed by quantitative real-time PCR, RNA-seq, and Western blotting. After chilling treatment, the transgenic lines showed significantly improved phenotype, higher kernel production, survival rate and biomass, and lower relative electrolyte leakage and superoxide dismutation than the untransformed line. Thus, ectopic expression of AnAFP gene improved chilling tolerance in the transgenic lines, which could be used to apply for further safety assessment for commercial breeding.

  • Research Papers
    Yu Wei, Liangjing Cao, Xucheng Huang, Xuan Wang, Huan Wang, Yongchao Song, Qiang He, Mingjie Lyu, Xinwen Hu, Jun Liu
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    The diploid wild goat grass Aegilops tauschii (Ae.?tauschii, 2n = 14; DD), as the D-sub genome of common wheat, provides rich germplasm resources for many aspects of wheat breeding. Abscisic acid (ABA) is an essential phytohormone that plays a pivotal role in plant adaptation to abiotic stresses. However, the gene regulation network of Ae.?tauschii in response to ABA stress remains unclear. Here, we conducted a time-course strand-specific RNA-sequencing study to globally profile the transcriptome that responded to ABA treatment in Ae.?tauschii. We identified 4818 differentially expressed transcription units/genes with time-point-specific induction/repression patterns. Using functional annotation, one-to-one ortholog and comparative transcriptome profiling analyses, we identified 319 ABA-responsive Ae.?tauschii orthologs that were also induced/repressed under ABA treatment in hexaploid wheat. On the quantitative trait loci (QTL) used in wheat marker-assisted breeding, we found that the ABA-responsive expression patterns of eight Ae.?tauschii orthologs were associated with drought stress tolerance, flowering process and/or grain quality. Of them, the ABA-responsive gene encoding sucrose:sucrose 1-fructosyltransferase in fructan and glucose metabolism pathways showed the most significant association with wheat drought tolerance. The characterization of ABA early-responsive genes in this study provides valuable information for exploring the molecular functions of the regulatory genes and will assist in wheat breeding.

  • Short Communication
    Ricardo St Aime, Grace Rhodes, Michael Jones, B. Todd Campbell, Sruthi Narayanan
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    Cotton (Gossypium spp.) is an important fiber and oil crop grown worldwide. Water and nutrient stresses are major issues affecting cotton production globally. Root traits are critical in improving water and nutrient uptake and maintaining plant productivity under optimal as well as drought conditions. However, root traits have rarely been utilized in cotton breeding programs, a major reason being the lack of information regarding genetic variability for root traits. The objective of this research was to evaluate ten selected cotton genotypes for root traits and water use efficiency. The tested genotypes included germplasm lines (PD 1 and PD 695) and cultivars that are currently grown in the southeastern USA (PHY 499WRF, PHY 444WRF, PHY 430W3FE, DP 1646B2XF, DP 1538B2XF, DP 1851B3XF, NG 5007B2XF, and ST 5020GLT). Experiments were conducted under controlled environmental conditions in 2018 and 2019. A hardpan treatment was included in the second year to evaluate the effect of a soil hardpan on root traits and water use efficiency. Genotype PHY 499WRF ranked at the top and NG 5007B2XF ranked at the bottom for root morphological traits (total and fine root length, surface area, and volume) and root weight. PHY 499WRF was also one of the best biomass producers and had high water use efficiency. PHY 444WRF, PHY 430W3FE, and PD-1 were the other best genotypes in terms of root traits and water use efficiency. All genotypes had higher values for root traits and water use efficiency under hardpan conditions. This trend indicates a horizontal proliferation of root systems when they incur a stress imposed by a hardpan. The genotypic differences identified in this research for root traits and water use efficiency would be valuable for selecting genotypes for cotton breeding programs.

  • Short Communication
    Huanhe Wei, Tianyao Meng, Jialin Ge, Xubin Zhang, Tianyi Shi, Enhao Ding, Yu Lu, Xinyue Li, Yuan Tao, Yinglong Chen, Min Li, Qigen Dai
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    Dense planting could be a feasible method for reducing nitrogen (N) application rates without compromising rice grain yield in northeast and central China. It is still unclear whether reduced N application with dense planting (RNDP) can achieve higher rice yield and N use efficiency (NUE) in Jiangsu, east China. Three japonica inbred rice (JI) and three indica hybrid rice (IH) cultivars were grown in a field experiment. Their grain yield, NUE, and related traits were compared under two cultivation treatments: conventional high-yielding practice (CHYP) and RNDP. JI showed similar yields under the two treatments, while IH showed lower yield under RNDP than under CHYP, and the partial factor productivity of N and N use efficiency for grain yield increased (P < 0.05) in both JI and IH under RNDP. Compared with CHYP, RNDP reduced spikelets per panicle but increased panicles per m2 and filled-kernel percentage of JI and IH, and JI’s kernel weight was increased (P < 0.05) under RNDP. Shoot biomass weight and nonstructural carbohydrate (NSC) content in the stem at heading and maturity of JI and IH were reduced under RNDP, while harvest index and NSC remobilization reserve were increased (P < 0.05) under RNDP, especially for JI. Our results suggest that RNDP could achieve a higher rice grain yield and NUE, particularly for JI, a dominant rice cultivar type in Jiangsu. For JI, the increased panicles per m2,the increased panicles per m2 < 0.05) under RNDP, especially for JI. Our results suggest that RNDP could achieve a higher rice grain yield and NUE, particularly for JI, a dominant rice cultivar type in Jiangsu. For JI, the increased panicles per m2, sink-filling efficiency, harvest index, and NSC remobilization after heading under RNDP contributed to a grain yield similar to that under CHYP.