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Co-silencing E1 and its homologs in an extremely late-maturing soybean cultivar confers super-early maturity and adaptation to high-latitude short-season regions
LIU Li-feng, GAO Le, ZHANG Li-xin, CAI Yu-peng, SONG Wen-wen, CHEN Li, YUAN Shan, WU Ting-ting, JIANG Bing-jun, SUN Shi, WU Cun-xiang, HOU Wen-sheng, HAN Tian-fu
2022, 21 (2): 326-335.   DOI: 10.1016/S2095-3119(20)63391-3
Abstract229)      PDF in ScienceDirect      
Soybean (Glycine max (L.) Merr.), a typical short-day plant, is sensitive to photoperiod, which limits the geographical range for its cultivation.  In the flowering pathway regulated by photoperiod, E1, a flowering inhibitor in soybean, plays the dominant role in flowering time regulation.  Two E1 homologs, E1-like-a (E1La) and E1-like-b (E1Lb), play overlapping or redundant roles in conjunction with E1.  In the present study, E1 and E1La/b were simultaneously silenced via RNA interference (RNAi) in Zigongdongdou (ZGDD), an extremely late-flowering soybean landrace from southern China.  As a result, RNAi lines showed a much earlier-flowering phenotype and obvious photoperiod insensitivity compared with wild-type (WT) plants.  In RNAi transgenic plants, the expression levels of flowering inhibitor GmFT4 and flowering promoters GmFT2a/GmFT5a were significantly down- and up-regulated, respectively.  Further, the maturity group (MG) of the RNAi lines was reduced from WT ZGDD’s MG VIII (extremely late-maturity) to MG 000 (super-early maturity), which can even grow in the northernmost village of China located at a latitude of 53.5°N.  Our study confirms that E1 and E1La/b can negatively regulate flowering time in soybean.  The RNAi lines generated in this study, with early flowering and maturity traits, can serve as valuable materials and a technical foundation for breeding soybeans that are adapted to high-latitude short-season regions.
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GmNMH7, a MADS-box transcription factor, inhibits root development and nodulation of soybean (Glycine max [L.] Merr.)
MA Wen-ya, LIU Wei, HOU Wen-sheng, SUN Shi, JIANG Bing-jun, HAN Tian-fu, FENG Yong-jun, WU Cun-xiang
2019, 18 (3): 553-562.   DOI: 10.1016/S2095-3119(18)61992-6
Abstract266)      PDF (1031KB)(220)      
As an important food crop and oil crop, soybean (Glycine max [L.] Merr.) is capable of nitrogen-fixing by root nodule.  Previous studies showed that GmNMH7, a transcription factor of MADS-box family, is associated with nodule development, but its specific function remained unknown.  In this study, we found that GmNMH7 was specifically expressed in root and nodule and the expression pattern of GmNMH7 was similar to several genes involved in early development of nodule (GmENOD40-1, GmENOD40-2, GmNFR1a, GmNFR5a, and GmNIN) after rhizobia inoculation.  The earlier expression peak of GmNMH7 compared to the other genes (GmENOD40-1, GmENOD40-2, GmNFR1a, GmNFR5a, and GmNIN) indicated that the gene is related to the nod factor (NF) signaling pathway and functions at the early development of nodule.  Over-expression of GmNMH7 in hairy roots significantly reduced the nodule number and the root length.  In the transgenic hairy roots, over-expression of GmNMH7 significantly down-regulated the expression levels of GmENOD40-1, GmENOD40-2, and GmNFR5α.  Moreover, the expression of GmNMH7 could respond to abscisic acid (ABA) and gibberellin (GA3) treatment in the root of Zigongdongdou seedlings.  Over-expressing GmNMH7 gene reduced the content of ABA, and increased the content of GA3 in the positive transgenic hairy roots.  Therefore, we concluded that GmNMH7 might participate in the NF signaling pathway and negatively regulate nodulation probably through regulating the content of GA3.
 
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Constitutive expression of feedback-insensitive cystathionine γ-synthase increases methionine levels in soybean leaves and seeds
YU Yang, HOU Wen-sheng, Yael Hacham, SUN Shi, WU Cun-xiang, Ifat Matityahu, SONG Shi-kui, Rachel Amir, HAN Tian-fu
2018, 17 (01): 54-62.   DOI: 10.1016/S2095-3119(16)61599-X
Abstract714)      PDF in ScienceDirect      
Soybean (Glycine max (L.) Merr.) is a major crop that provides plant-origin protein and oil for humans and livestock.  Although the soybean vegetative tissues and seeds provide a major source of high-quality protein, they suffer from low concentration of an essential sulfur-containing amino acid, methionine, which significantly limits their nutritional quality.  The level of methionine is mainly controlled by the first unique enzyme of methionine synthesis, cystathione γ-synthase (CGS).  Aiming to elevate methionine level in vegetative tissues and seeds, we constitutively over-expressed a feedback-insensitive
Arabidopsis CGS (AtD-CGS) in soybean cultivars, Zigongdongdou (ZD) and Jilinxiaoli 1 (JX).  The levels of soluble methionine increased remarkably in leaves of transgenic soybeans compared to wild-type plants (6.6- and 7.3-fold in two transgenic ZD lines, and 3.7-fold in one transgenic JX line).  Furthermore, the total methionine contents were significantly increased in seeds of the transgenic ZD lines (1.5- to 4.8-fold increase) and the transgenic JX lines (1.3- to 2.3-fold increase) than in the wild type.  The protein contents of the transgenic soybean seeds were significantly elevated compared to the wild type, suggesting that the scarcity of methionine in soybeans may limit protein accumulation in soybean seeds.  The increased protein content did not alter the profile of major storage proteins in the seeds.  Generally, this study provides a promising strategy to increase the levels of methionine and protein in soybean through the breeding programs.  
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Analysis of the independent- and interactive-photo-thermal effects on soybean flowering
WU Ting-ting, LI Jin-yu, WU Cun-xiang, SUN Shi, MAO Ting-ting, JIANG Bing-jun, HOU Wen-sheng, HAN Tian-fu
2015, 14 (4): 622-632.   DOI: 10.1016/S2095-3119(14)60856-X
Abstract2049)      PDF in ScienceDirect      
Soybean (Glycine max (L.) Merr.) is a typical short-day and warm season plant, and the interval between emergence and flowering has long been known to be regulated by environmental factors, primarily photoperiod and temperature. While the effects of photoperiod and temperature on soybean flowering have been extensively studied, a dissection of the component photo-thermal effects has not been documented for Chinese germplasm. Our objective of the current study was to evaluate the independent- and interactive-photo-thermal responses of 71 cultivars from 6 ecotypes spanning the soybean production regions in China. These cultivars were subjected in pot experiments to different temperature regimes by planting in spring (low temperature (LT)) and summer (high temperature (HT)), and integrating with short day (SD, 12 h), natural day (ND, variable day-length), and long day (LD, 16 h) treatments over two years. The duration of the vegetative phase from emergence to first bloom (R1) was recorded, and the photo-thermal response was calculated. The outcome of this characterization led to the following conclusions: (1) There were significant differences in photo-thermal response among the different ecotypes. High-latitude ecotypes were less sensitive to the independent- and interactive-photo-thermal effects than low-latitude ecotypes; and (2) there was an interaction between photoperiod and temperature, with the effect of photoperiod on thermal sensitivity being greater under the LD than the SD condition, and with the effect of temperature on photoperiodic sensitivity being greater under the LT than the HT condition. The strengths and limitations of this study are discussed in terms of implications for current knowledge and future research directions. The study provides better understanding of photo-thermal effects on flowering in soybean genotypes from different ecotypes throughout China and of the implications for their adaptation more broadly.
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Construction of Two Suppression Subtractive Hybridization Libraries and Identification of Salt-Induced Genes in Soybean
LI Liang, WANG Wei-qi, HAN Tian-fu, HOU Wen-sheng
2012, 12 (7): 1075-1085.   DOI: 10.1016/S1671-2927(00)8632
Abstract1467)      PDF in ScienceDirect      
Soybean is planted worldwide and its productivity is significantly hampered by salinity. Development of salt tolerant cultivars is necessary for promoting soybean production. Despite wealth of information generated on salt tolerance mechanism, its basics still remain elusive. A continued effort is needed to understand the salt tolerance mechanism in soybean using suitable molecular tools. To better understand the molecular basis of the responses of soybean to salt stress and to get an enrichment of critical salt stress responsive genes in soybean, suppression subtractive hybridization libraries (SSH) are constructed for the root tissue of two cultivated soybean genotypes, one was tolerant and the other was sensitive to salt stress. To compare the responses of plants in salt treatment and non-treatment, SSH1 was constructed for the salt-tolerant cultivar Wenfeng 7 and SSH2 was constructed for the salt-sensitive cultivar Union. From the two SSH cDNA libraries, a total of 379 high quality ESTs were obtained. These ESTs were then annotated by performing sequence similarity searches against the NCBI nr (National Center for Biotechnology Information protein non-redundant) database using the BLASTX program. Sixty-three genes from SSH1 and 49 genes from SSH2 could be assigned putative function. On the other hand, 25 ESTs of SSH1 which may be not the salt tolerance-related genes were removed by comparing and analyzing the ESTs from the two SSH libraries, which increased the proportion of the genes related to salt tolerance in SSH1. These results suggested that the novel way could realize low background of SSH and high level enrichment of target cDNAs to some extent.
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