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Article

Effects of Salinity-Stress on Seed Germination and Growth Physiology of Quinclorac-Resistant Echinochloa crus-galli (L.) Beauv

1
Hunan Weed Science Key Laboratory, Hunan Academy of Agricultural Sciences, Changsha 410125, China
2
Hunan Agricultural Biotechnology Research Institute, Hunan Academy of Agricultural Sciences, Changsha 410125, China
*
Author to whom correspondence should be addressed.
Submission received: 13 April 2022 / Revised: 10 May 2022 / Accepted: 11 May 2022 / Published: 15 May 2022
(This article belongs to the Special Issue Herbicides Toxicology and Weeds Herbicide-Resistant Mechanism)

Abstract

:
With the expansion of saline-alkaline tolerant rice in China, the effects of salinity stress (NaCl) on quinclorac-resistant Echinochloa crus-galli (L.) Beauv (E. crus-galli) is unclear. In this study, the growth chamber experiment was conducted to test the germination and growth physiology of seven populations of E. crus-galli with quinclorac-different resistance levels which were collected from Hunan province. The results showed that a significant decrease of the germination rate and fresh weight of the plants, as well as the length of the roots and young shoots appeared, along with the increased resistance while treated with NaCl. However, no significant differences were detected between quinclorac-resistant and -susceptible E. crus-galli populations while without NaCl treatment. A further study with spectrophotometer showed that the salinity treatment resulted in the increase of the GST activity in all E. crus-galli populations, which are more obvious in those resistant biotypes, and transcriptomics revealed that salt stress reduces the adaptability of quinclorac-resistant E. crus-galli by reducing the biosynthesis, activities of antioxidant enzymes and metabolic enzyme. This study demonstrated that salinity stress (NaCl) may reduce the adaptability of quinclorac-resistant E. crus-galli.

1. Introduction

Millions of people feed on rice, which makes rice a vital cereal crop in the world. To feed the increasing number of people, a higher productivity of rice in limited acreage was pursued at the cost of the ecosystem, which in turn adversely affects the production of rice. For example, soil salinity brings lower yield of the grain [1], and is becoming a primary abiotic constraint against the production of crops globally. The salinity adversely damages multiple important physiological processes, such as germination and seedling growth [2]. Previous studies showed that the side effects of salinity on both germination and seedling growth arose from the external osmotic potential which reduces the uptake of water, or from the toxic effect of Cl and Na+ ions on the germinating seed [3]. Soil with an electrical conductivity of the saturation soil extracts of more than 4 dS/m (corresponding to about 40 mM NaCl) at 25 °C is categorized as saline [4,5].
Besides salinity stress, weeds, which are life-long companions of rice, can also reduce rice yield and quality. At a density of 9 plants m−2, the presence of the primary weed, E. crus-galli, costs more than 50% reduction of the rice [6,7]. A heavy infection of such weed can remove up to 80% of the nitrogen (N) from the soil [8]. The quinclorac, one of the auxinic herbicides to control E. crus-galli in rice fields, has been widely used in rice fields of China since the 1990s [9]. Nowadays, although the rapid development of herbicide resistance in E. crus-galli is developing [10,11,12], a better adaptability of the weed to the extreme environment is reported [13,14], and the adverse influence on seed germination of E. crus-galli was observed [15].
The activity and expression of glutathione S-transferases (GST) of plants was reported as two more factors against the salinity. For example, a cDNA encoding an enzyme with both glutathione peroxidase (GPX) and glutathione S-transferase activity had GPX- specific activities in the transgenic tobacco seedlings was over-expressed up to two-times that in the wild-type. Under the condition of chilling and salt stress, the over expression of GST/GPX stimulates the growth of the seedling [16]. However, few studies on the effect and mechanism of salt stress on the growth of herbicides-resistant weeds were conducted. Here, this study aims to (1) reveal the effect of salt stress on seed germination and seedling growth of quinclorac—E. crus-galli; (2) clarify the changing regularities of GST activities under salt stress; and (3) how salt stress affects the physiology of quinclorac-resistant E. crus-galli. The findings can promote weed management in rice fields under saline environment.

2. Materials and Methods

2.1. Plant Materials

The experiments were conducted at Hunan Agricultural Biotechnology Research Institute, Changsha, Hunan. The seeds of E. crus-galli were collected from two adjacent rice fields in Chunhua (field X and field Y), Hunan Province in 2013. The failure to control E. crus-galli with an application of quinclorac for 15 years was reported in field X (28.31° N, 113.30° E), while no quinclorac was sprayed in field Y (28.29° N, 113.27° E) during the previous five years. The seeds of E. crus-galli from field X and Y were collected and cultivated to tillering in plastic pots (30 cm × 20 cm × 12 cm) in a greenhouse. Two tillers were collected from each plant, with one for seed collection and the other for sensitivity analysis. The sensitivity was determined by the treatment of the tiller with quinclorac (400 g a.i./ha, 96%, Rainbow Shandong Province, PD20142361). All biotype Y E. crus-galli plants died at 21 days after the treatment of quinclorac; whilst the X counterparts were all alive. The dead tillers were identified as susceptible, while the surviving tillers classified as resistant. The resistant and susceptible seeds were all preserved [17].
The seeds were propagated for four generations at a breeding pool from 2014 to 2017. The quinclorac-susceptible biotype was named QS, while the quinclorac-resistant biotype was named QR. The other five E. crus-galli populations were collected in 2017 from Dingcheng (DC, 28.92° N, 111.76° E), Hanshou (HS, 29.11° N, 111.89° E), Shimen (SM, 29.64° N, 111.20° E), Hekou (HK, 27.71° N, 112.89° E) and Chunhua (CH, 28.31° N, 113.29° E). All of these populations had been sprayed with quinclorac more than once.

2.2. Detection of Quinclorac-Resistance Level of All Selected E. crus-galli Populations

Healthy and full seeds of E. crus-galli were selected and soaked for 24 h in 0.5% KNO3 for dormancy-breaking. Then, the seeds were washed and sown in a plastic pot (6 cm in height and 10 cm in diameter). To accelerate germination, the seeds were cultivated at 28 °C. When the seedlings reached 3–4 leaf stage, the quinclorac was sprayed using a spray tower (3WP-2000, Nanjing Research Institute for Agricultural Mechanization Ministry of Agriculture, Nanjing, China), at a density of 0, 200, 400, 800, 1600 and 3200 g a.i./ha to the QR and CH biotype, and at a density of 12.5, 25, 50, 100 and 200 g a.i./ha to the other five populations. All the above parts of the plants were cut to weigh the weight of each pot after three weeks’ treatment. All the experiments were performed with three replications in a completely randomized design. The rate of fresh weight inhibition and relative resistance index was the same as mentioned by Wu, et al. (2019) [17].

2.3. Effects of Salinity Stress on the Seed Germination of E. crus-galli

Five concentrations of NaCl aqueous with 0, 20, 50, 100 and 150 mM were used for the simulation of salinity stress. To verify how salinity affects the germination of E. crus-galli, the full and healthy seeds which were dormancy-broken with the method described above, were air dried. The dry seeds (quantity 100) were placed in a Petri dish (9-cm-diameter) containing two layers of Whatman filter paper moistened with 10 mL sodium chloride (NaCl) aqueous or deionized water [18]. To minimize evaporation, the Petri dish made of 3-mm-thick glass, was properly sealed by parafilm. The germination rate of the seed was investigated after 48, 72 and 96 h post-treatment.

2.4. Effects of Salinity Stress on the Young Shoots and Roots Growth of E. crus-galli

The seeds were treated as above with the five concentrations of NaCl solution. All twenty germinated seeds were cultivated at 28 °C. The length of the young shoots and roots of the various treatments were measured. Each experiment was conducted in triplicate.

2.5. Effects of Salinity Stress on the Fresh Weight of E. crus-galli

The young 2–3 cm tall buds of E. crus-galli were chosen and treated with the various concentrations of NaCl solution. The various concentrations of NaCl solutions were obtained by dissolving in the Hoagland’s solution, with final concentrations of 0, 20, 50, 100 and 150 mM. The plants treated with NaCl solution were raised in a 4-L rectangular plastic box (25 cm × 16 cm × 10 cm) with 3.5 L of Hoagland’s solution (containing NaCl). Foam plates having 20 small holes (10 by 2 arrangements) with gauze were placed at the bottoms of the rectangular plastic box [14]. The seedlings were transplanted into the holes, with two plants in each hole. The fresh weight was investigated among the various treatments after 21 days treatments.

2.6. Effects of Salinity Stress on the GST Activity of E. crus-galli

The concentration of 50 mM NaCl was applied when the plants had grown to 3–4-leaves in Hoagland’s solution. The samples were respectively collected at 0, 1, 3, 5 and 14 days after treatment. The fresh leaves were removed and weighed, then wrapped in silver paper and finally put into liquid nitrogen, and stored in a refrigerator at −80 ℃. The leaf was homogenized in liquid nitrogen. The homogenate was centrifuged at 12,000× g for 20 min and filtered. The GST assay kit (Nanjing Jiancheng Bioengineering Institute, Nanjing, China, A004) was used, and protein content was determined with the method previously published by Bradford (1976) [19]. GSH formation was recorded at 28 ℃ on a spectrophotometer 721 (INESA, Shanghai, China) at 412 nm.

2.7. Transcriptomics of Quinclorac-Resistant and Sensitive E. crus-galli under Salt Stress

First, total RNA was extracted from young leaf tissues of the 3–4 leaf stage E. crus-galli (QR and QS) which were treated with 50 mM NaCl for 3 d using TransZol Up Plus RNA Kit (TransGen Biotech Co., Ltd., Beijing, China), and the E. crus-galli without NaCl treatment was the control; the whole experiment was repeated three times. Second, mRNA was enriched by magnetic beads using Dynabeads Oligo (dT)25 (Thermo Fisher, Waltham, MA, USA, 61002), then 10× fragmentation buffer (Enzymatics, Beverly, MA, USA, B0330) was added to the purified mRNA to break its fragments into short fragments. Third, the first and second strands of cDNA were synthesized with the fragmented mRNA as the template, then the library was constructed from the synthesized double stranded cDNA. Finally, the qualified library was sequenced on Illumina platform, and the sequencing strategy was PE 150. Unqualified sequencing reads were removed to obtain clean reads, and the clean reads were aligned to E. crus-galli_v6.prot and Oryza sativa subsp. Japonica reference genome. The gene expression levels were estimated via the FPKM (fragment per kilobase transcriptome per million mapped reads) method [20]. The differentially expressed genes (DEGs) between the QRNaCl, QSNaCl, QRCK and QSCK were analyzed with the DESeq R package (1.18.0), and the genes with an adjusted p-value < 0.05 were classified as DEGs.

2.8. Statistical Analysis

All experiments were conducted in a completely randomized design with three replications, and each experiment was conducted twice. The data were the mean of two runs, and there was no significant difference between both experimental runs. Data were submitted to analysis of variance, the averages of the germination rate, the shoot length, the root length and the fresh weight were compared by the Duncan’s multiple-range test at 5% significance, all data were calculated using SPSS 10.0. The figures were drawn using Sigma Plot 10.0.

3. Results

3.1. The Susceptibility of E. crus-galli to Quinclorac

Significant difference in quinclorac resistance was observed among the seven selected E. crus-galli populations (Table 1). Compared to the QS biotype with a GR50 of 27.65 g a.i./ha, the CH and QR populations were relatively unsusceptible to quinclorac, whilst the GR50 of CH and QR (4417.91 g. a.i./ha and 3855.01 g. a.i./ha, respectively) were much higher than the recommended field dose (225–375 g. a.i./ha, China). Based on the resistance index (RI), CH populations had GR50 values 159.78-fold higher than that of QS, while that of the CH biotype was 139.42-fold higher; the relative RI values of the HS, SM, HK and DC populations were 5.12, 3.13, 2.32 and 1.14, respectively. Therefore, the highest resistance among the seven E. crus-galli populations was CH, following by QR, HS, SM, HK, DC and QS populations.

3.2. Seed Germination of Seven E. crus-galli Populations under Salinity Stress

The salinity stress inhibited the germination of the E. crus-galli seeds, and a negative relation between germination rate of the seed and the corresponding resistance of E. crus-galli was concluded. In addition, the germination rate at 96 h (Figure 1c) was significantly higher than that at 72 h (Figure 1b) and 48 h (Figure 1a) post treatment, while there were no significant differences in the resistance levels in the plants without NaCl treatment (Tables S1–S3).

3.3. Shoots and Roots Growth of Seven E. crus-galli Populations under Salinity Stress

In this study, similar reduction in the lengths of shoots (Figure 2, Tables S4 and S5) and roots (Figure 3, Tables S6 and S7) of E. crus-galli was also observed, when treated with salinity. The higher the concentration of NaCl, the greater the inhibitory effect on shoots and roots of E. crus-galli. The length at 7 days (Figure 2b and Figure 3b) was higher than that at 4 days post treatment (Figure 2a and Figure 3a). With increasing quinclorac-resistant level, the growth of young shoots and roots of E. crus-galli was obviously affected by the NaCl solution. When 20 mM NaCl was applied, positive effects on the shoots and roots growth of seven populations were detected, while a severe inhibition was observed with 150 mM NaCl, especially for the QR biotype, which was one of the types most resistant to quinclorac. There were no significant differences in the growth of the shoots and roots among the seven populations without NaCl treatment.

3.4. Fresh Weight of Seven E. crus-galli Populations under Salinity Stress

The salinity-stress has a significant inhibitory effect on fresh weight of E. crus-galli. In this study, salinity decreased the fresh weight of seven E. crus-galli populations to a different extent. Compared with the sensitive populations, a more severe inhibition in the fresh weight of E. crus-galli was observed in higher quinclorac-resistance level. When the concentration of NaCl was 150 mM (Figure 4), the fresh weight of CH and QR was 0.18 g (10 plants), while the other five populations were 0.2–0.51 g (10 plants), (Table S8).

3.5. GST Activity Analysis

In the present study, enzymatic activity was determined with the sprouts of E. crus-galli using a GST assay kit, based on spectrophotometer with the general GST substrate CDNB. The CDNB was used to detect GST in all organisms [21]. The results showed that the QS populations, the most sensitive to quinclorac among the seven types, had the highest GST activity under 50 mM NaCl treatment. When treated for 3 days, the GST activity reached 374 U/mgprot. The CH biotype with the strongest resistant to quinclorac in this study, had the lowest GST activity. The GST activity of the seven E. crus-galli populations ranging from low to high were as follows: CH, QR, SH, SM, HK, DC, and QS. The trends of the GST activity of all seven populations were an initial increase followed by a subsequent decrease (Figure 5a, Table S9). While without NaCl-stress, a negative relation between the quinclorac-resistance level of E. crus-galli and GST was observed along with time, however there were no significant changes in GST activity among the seven populations, respectively (Figure 5b, Table S10).

3.6. Transcription and Functional Analysis of E. crus-galli Induced by Salinity Stress

All of the DEGs were assigned and classified into three groups (biological process, cellular component and molecular function). When under NaCl stress, within the three main categories, the largest proportion of the biological process category was represented by metabolic process (39.80%) and cellular process (37.70%), compared with the control group, 656 and 581 related genes were increased, respectively; in the cellular component category, it was cell part (48.93%) and organelle (27.43%), 408 and 771 related genes were increased, respectively, compared with the control group; and in the molecular function category, it was catalytic activity (41.21%) and binding (36.51%), which were 685 and 574 related genes were increased, respectively, compared with the control group (Figure 6 and Figure 7, Tables S11 and S12).
Following NaCl treatment in the QR and QS biotypes of E. crus-galli, 563 DEGs in the QR samples relative to the QS samples were annotated into KEGG pathways. Pathway enrichment analysis found that the most highly enriched pathways included the “linoleate 9S-lipoxygenase” (K15718), “solute carrier family” (K14709, K14445), “arogenate/prephenate dehydratase” (K05359), “monodehydroascorbate reductase” (K08232), “Cytochrome P450” (K05280), “peroxidase” (K00430), “cellulose synthase A” (K10999) and “glutathione S-transferase” (K00799)_pathways. Four herbicide-related species, such as “Cytochrome P450”, “glutathione S-transferase”, were found to be down-accumulated. Seven DEGs associated with “defense response” were found responsive to salt stress, which is involved in the regulation of stress processes.

4. Discussion

Salinity is one of the most serious environmental issues in the world and has a negative impact on crop production [22,23]. Karimi et al. (2018) found that Banebaghi fresh and dry weight of shoot and root in both stem height groups decreased with increasing levels of salinity [24]. Salinity and other adversities, such as salicylic acid, also affect the herbicide tolerance of the plants by regulating physiological and biochemical reaction [25,26]. For instance, to protect the rice, SA enhanced the expression of some genes to accelerate the metabolism of quinclorac under quinclorac stress [27]. Salinization not only reduces the yield and quality of crops [28], but is also one of the major and widespread challenges hindering global food security and environmental sustainability [29]. Is salinity not beneficial for crops? While compared with no irrigation, saline water application can increase crop yields but may lead to some adverse effects on soil properties and plant growth. Water salinity less than 3.2 dS m−1 stimulated root growth in the soil layers 10–40 cm at the heading stage without obvious yield reduction or salt accumulation, and the interaction effects of irrigation amount and water salinity had no statistically significant (p > 0.05) effects on root parameters and yield [30].
Salinity stress also affects the germination and growth of weeds. When treated with 50 mM NaCl, the seed germination of Echinochloa colona was larger than 60% and some seeds still germinated at 150 mM NaCl [31]. Additionally, germination capacity of the sensitive E. crus-galli was higher (up to 90%) than that of the ALS-inhibitor herbicides resistant populations (about 70%) while NaCl concentration was from 0 to 250 mM, and, speed of germination and root and shoot length of seedlings were also inversely related to salt concentration [32]. In China, quinclorac-resistant E. crus-galli was first discovered in Hunan Province in 2002 [33], then further discovery of some other resistant populations [34,35], However, there are few reports on the effects of salt stress on quinclorac-resistant E. crus-galli. This study found that the germination of quinclorac-resistant E. crus-galli populations are more sensitive to salinity stress. In the CH biotype, the resistance level of quinclorac is the highest, which may result from the constant use of quinclorac application for 15 years. Additionally, the different quinclorac resistances of HS, SM, HK and DC populations may also be due to the varying levels of quinclorac applied. The salinity stress significantly inhibited the quinclorac-resistant E. crus-galli germination, and the higher the resistance, the lower the germination rate.
Salinity adversely affects plant growth, such as the reduction of the crop yield by altering plant metabolism, including reduced water potential, ion imbalances and toxicity, even a whole crop failure [36]. Different plants have different tolerances to salt stress, i.e., compared with cowpea, cotton is more tolerant to salt [37]. Noreen et al. (2013) studied the effects of salinity on the length of the root and shoot of moringa plants, and found that the highest reduction in root and shoot length was at 20 dS m−1 [38]. The growth of weed species can also be affected by the salinity. Our study found that the shoot length, root length and fresh weight of quinclorac-resistant E. crus-galli were all inhibited significantly by salinity stress compared with the sensitive populations. The lower germination rate of the weeds and slower growth rate of roots and shoots in the higher quinclorac-resistant levels of E. crus-galli indicated that the quinclorac-resistant weeds cost a lot under drought stress, similar with the findings of Kukorelli, et al. [39,40,41].
The expression of GSTs in plants is regarded as highly responsive to biotic and abiotic stresses, a wide variety of stress-associated chemicals and other synthetic and natural auxins [42]. The GSTs are also best known for their ability to conjugate GSH to a number of herbicides and thus reduce their toxicity [43]. Thus, the GSTs are considered to play an important role in plant salt tolerance [44]. This work showed that the salinity stress significantly inhibited the GST activity of quinclorac-resistant E. crus-galli, which may lead to poorer adaptability of quinclorac-resistant E. crus-galli to salinity. Based on the role of GST in plant salt tolerance, we speculate that sensitive populations promoted the adaptability of salinity accompanying a rise in GST activity. By reducing GST activity in quinclorac-resistant E. crus-galli, the control efficiency of resistant weeds should be improved. Furthermore, transcriptomics revealed that the expression of related genes, such as cytochrome P450 and peroxidase (POD) in resistant E. crus-galli were downregulated while treated with NaCl, indicating that resistant E. crus-galli was weaker in photosynthesis and redox reaction than sensitive ones. This was consistent with the previous report that the plant height and fresh weight of resistant E. crus-galli under salt stress were lower than that of sensitive E. crus-galli, indicating that the adaptability of resistant E. crus-galli to salt environment is lower than that of sensitive biotypes, which is the fitness cost paid by resistant E. crus-galli to adapt to adverse stress.
Our findings revealed that quinclorac-resistance in E. crus-galli reduced its adaptability to salinity, and with the exception of applying other herbicides, the regulating of the soil may represent a good method for managing the quinclorac resistance of E. crus-galli. Therefore, these data are valuable for increasing agricultural production and the incomes of the farmers. Or through formulation processing, such as cationic covalent organic frameworks and other methods to improve the efficacy of herbicides, so as to achieve the purpose of controlling resistant weeds [45].

5. Conclusions

Overall, we have identified the effects of salt stress on germination, seedling growth and their GST activity of E. crus-galli populations with different quinclorac resistance levels. While under salt stress, the higher level of quinclorac-resistance leading to the greater inhibitory effect on seed germination, shoots and roots length and fresh weight of E. crus-galli, but the lower GST activity. Besides, through transcriptome sequencing, we found many DEGs between resistance and sensitive E. crus-galli, which regulate the response to salt stress. We will carry out functional verification for key genes in the future, which will lay a foundation in the management of the resistant weeds.

Supplementary Materials

The following supporting information can be downloaded at: https://0-www-mdpi-com.brum.beds.ac.uk/article/10.3390/agronomy12051193/s1, Table S1: The germination of the seven E. crus-galli populations treated by salinity stress for 48 h; Table S2: The germination of the seven E. crus-galli populations treated by salinity stress for 72 h; Table S3: The germination of the seven E. crus-galli populations treated by salinity stress for 96 h; Table S4: The shoot length of seven E. crus-galli populations treated by salinity stress for 4 days; Table S5: The shoot length of seven E. crus-galli populations treated by salinity stress for 7 days; Table S6: The root length of seven E. crus-galli populations treated by salinity stress for 4 days; Table S7:The root length of seven E. crus-galli populations treated by salinity stress for 7 days; Table S8: The fresh weight of seven E. crus-galli populations treated by salinity stress for 7 days; Table S9: The GST activity of E. crus-galli under 50 mM NaCl treatment at different times; Table S10: The GST activity of E. crus-galli without NaCl at different times; Table S11: The Gene Ontology (GO) analysis of the DEGs between QR and QS E. crus-galli populations without NaCl; Table S12: The Gene Ontology (GO) analysis of the DEGs between QR and QS E. crus-galli populations treated with 50 mM NaCl.

Author Contributions

Conceptualization, L.W. (Lamei Wu); methodology, L.W. (Lamei Wu); software, L.W. (Lamei Wu); validation, H.Y., Z.L. and Q.P.; resources, L.W. (Lamei Wu); data curation, L.W. (Lifeng Wang); writing—original draft preparation, L.W. (Lamei Wu); writing—review and editing, L.W. (Lamei Wu) and H.Y.; project administration, L.W. (Lamei Wu); funding acquisition, L.W. (Lamei Wu) and L.W. (Lifeng Wang). All authors have read and agreed to the published version of the manuscript.

Funding

This research was funded by the Science and Technology Innovation Program of Hunan Province project (No. 2020WK2023), the China Agriculture Research System of MOF and MARA project (No. CARS-16-E19), Hunan Provincial Key Laboratory for Biology and Control of Weed (No. 2015TP1016).

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

Not applicable.

Acknowledgments

We are grateful to many farmers for the provision of E. crus-galli seed samples. We also thank Xiangying Liu, Deng Xile, Lang Pan and anonymous reviewers for comments on the article.

Conflicts of Interest

The authors declare no conflict of interest.

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Figure 1. The germination rate (%) of the seven E. crus-galli populations at different time points under salinity stress. The germination rate (%) of E. crus-galli under four NaCl concentrations at (a) 48 h, (b) 72 h and (c) 96 h after treatment. CH, the E. crus-galli biotype collected from Chunhua, Hunan province, China; QR, the E. crus-galli biotype collected from Chunhua, Hunan province, China, which were bred for four generations from 2014 to 2017, and resistant to quinclorac; HS, the E. crus-galli biotype collected from Hanshou, Hunan province, China; SM, the E. crus-galli biotype collected from Shimen, Hunan province, China; HK, the E. crus-galli biotype collected from Hekou, Hunan province, China; DC, the E. crus-galli biotype collected from Dingcheng, Hunan province, China; QS, the E. crus-galli biotype collected from Chunhua, Hunan province, China, which were bred for four generations from 2014 to 2017, and sensitive to quinclorac. The same as below. Lowercase letters are expressed as least square means ± standard error (p < 0.05).
Figure 1. The germination rate (%) of the seven E. crus-galli populations at different time points under salinity stress. The germination rate (%) of E. crus-galli under four NaCl concentrations at (a) 48 h, (b) 72 h and (c) 96 h after treatment. CH, the E. crus-galli biotype collected from Chunhua, Hunan province, China; QR, the E. crus-galli biotype collected from Chunhua, Hunan province, China, which were bred for four generations from 2014 to 2017, and resistant to quinclorac; HS, the E. crus-galli biotype collected from Hanshou, Hunan province, China; SM, the E. crus-galli biotype collected from Shimen, Hunan province, China; HK, the E. crus-galli biotype collected from Hekou, Hunan province, China; DC, the E. crus-galli biotype collected from Dingcheng, Hunan province, China; QS, the E. crus-galli biotype collected from Chunhua, Hunan province, China, which were bred for four generations from 2014 to 2017, and sensitive to quinclorac. The same as below. Lowercase letters are expressed as least square means ± standard error (p < 0.05).
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Figure 2. The shoot length (cm) of E. crus-galli with four NaCl concentrations at different time points. The shoot length (cm) of E. crus-galli under four NaCl concentrations at 4 days (a) and 7 days (b) after treatment. Lowercase letters are expressed as least square means ± standard error (p < 0.05).
Figure 2. The shoot length (cm) of E. crus-galli with four NaCl concentrations at different time points. The shoot length (cm) of E. crus-galli under four NaCl concentrations at 4 days (a) and 7 days (b) after treatment. Lowercase letters are expressed as least square means ± standard error (p < 0.05).
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Figure 3. The root length (cm) of E. crus-galli with four NaCl concentrations at different time points. The root length (cm) of E. crus-galli under four NaCl concentrations at 4 days (a) and 7 days (b) after treatment. Lowercase letters are expressed as least square means ± standard error (p < 0.05).
Figure 3. The root length (cm) of E. crus-galli with four NaCl concentrations at different time points. The root length (cm) of E. crus-galli under four NaCl concentrations at 4 days (a) and 7 days (b) after treatment. Lowercase letters are expressed as least square means ± standard error (p < 0.05).
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Figure 4. The fresh weight (g) of E. crus-galli with four NaCl concentrations at 21 days after treatment. Lowercase letters are expressed as least square means ± standard error (p < 0.05).
Figure 4. The fresh weight (g) of E. crus-galli with four NaCl concentrations at 21 days after treatment. Lowercase letters are expressed as least square means ± standard error (p < 0.05).
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Figure 5. The GST activity of E. crus-galli under 50 mM NaCl treatment. The GST activity of E. crus-galli with 50 mM NaCl (a) and without NaCl treatment (b). Lowercase letters are expressed as least square means ± standard error (p < 0.05).
Figure 5. The GST activity of E. crus-galli under 50 mM NaCl treatment. The GST activity of E. crus-galli with 50 mM NaCl (a) and without NaCl treatment (b). Lowercase letters are expressed as least square means ± standard error (p < 0.05).
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Figure 6. Gene Ontology (GO) analysis of the DEGs between two E. crus-galli populations. GO analysis was performed at level 2 for the three main categories.
Figure 6. Gene Ontology (GO) analysis of the DEGs between two E. crus-galli populations. GO analysis was performed at level 2 for the three main categories.
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Figure 7. Gene Ontology (GO) analysis of the DEGs between two E. crus-galli populations treated with 50 mM NaCl. GO analysis was performed at level 2 for the three main categories.
Figure 7. Gene Ontology (GO) analysis of the DEGs between two E. crus-galli populations treated with 50 mM NaCl. GO analysis was performed at level 2 for the three main categories.
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Table 1. Quinclorac dose–response analyses for E. crus-galli populations.
Table 1. Quinclorac dose–response analyses for E. crus-galli populations.
E. crus-galli PopulationsGR50 (g a.i./ha)Resistant Index
CH4417.91159.78
QR3855.01139.42
HS141.645.12
SM86.643.13
HK64.122.32
DC31.461.14
QS27.651
Abbreviations: CH, the E. crus-galli biotype collected from Chunhua, Hunan province, China; QR, the E. crus-galli biotype collected from Chunhua, Hunan province, China, which were bred for four generations from 2014 to 2017, and resistant to quinclorac; HS, the E. crus-galli biotype collected from Hanshou, Hunan province, China; SM, the E. crus-galli biotype collected from Shimen, Hunan province, China; HK, the E. crus-galli biotype collected from Hekou, Hunan province, China; DC, the E. crus-galli biotype collected from Dingcheng, Hunan province, China; QS, the E. crus-galli biotype collected from Chunhua, Hunan province, China, which were bred for four generations from 2014 to 2017, and sensitive to quinclorac.
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Wu, L.; Yang, H.; Li, Z.; Wang, L.; Peng, Q. Effects of Salinity-Stress on Seed Germination and Growth Physiology of Quinclorac-Resistant Echinochloa crus-galli (L.) Beauv. Agronomy 2022, 12, 1193. https://0-doi-org.brum.beds.ac.uk/10.3390/agronomy12051193

AMA Style

Wu L, Yang H, Li Z, Wang L, Peng Q. Effects of Salinity-Stress on Seed Germination and Growth Physiology of Quinclorac-Resistant Echinochloa crus-galli (L.) Beauv. Agronomy. 2022; 12(5):1193. https://0-doi-org.brum.beds.ac.uk/10.3390/agronomy12051193

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Wu, Lamei, Haona Yang, Zuren Li, Lifeng Wang, and Qiong Peng. 2022. "Effects of Salinity-Stress on Seed Germination and Growth Physiology of Quinclorac-Resistant Echinochloa crus-galli (L.) Beauv" Agronomy 12, no. 5: 1193. https://0-doi-org.brum.beds.ac.uk/10.3390/agronomy12051193

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