Evaluation of saline-alkali tolerance in 71 soybean germplasms based on multi-method integrated analysis at the germination stage

Background Global soybean production is constrained by scarce arable land, and standardized evaluation tools remain lacking for natural mixed saline-alkali stress, the predominant abiotic stress under field conditions. Objective This study aimed to establish a comprehensive saline-alkali tolerance evaluation system for soybean germplasms via integrated multivariate statistical methods, and screen core and auxiliary indicators for efficient germplasm identification. Methods Seventy-one soybean germplasms were tested under 90 mmol/L mixed saline-alkali stress (NaCl:Na 2 SO 4 :NaHCO 3 :Na 2 CO 3 = 1:9:9:1, pH 8.2) simulating natural saline-alkali soil. We quantified the saline-alkali tolerance coefficients (SATC) of 13 morphological and physiological indicators, followed by coefficient of variation (CV), principal component analysis (PCA), subordinate function, cluster analysis and regression modeling. Results Significant inter-germplasm variations in saline-alkali tolerance were detected, and indicators with CV > 0.35 ( e.g ., root length (RL), root fresh weight (RFW)) were screened as primary indices. PCA extracted five principal components with 87.18% cumulative variance contribution, and the integrated analytical pipeline categorized germplasms into five tolerance grades: eight highly tolerant, 24 moderately tolerant, 13 generally tolerant, 16 sensitive and 10 highly sensitive accessions. A high-precision prediction model was constructed ( D = 0.290 X 1 − 0.026 X 2 + 0.438 X 3 + 0.402 X 4 + 0.180 X 5 + 0.153 X 6 + 0.813 X 7 − 1.123; R 2 = 0.998, where X 1 – X 7 represent the SATC of germination rate (GR), RL, RFW, total fresh weight (TFW), shoot dry weight (SDW), root dry weight (RDW), and total dry weight (TDW), respectively). A Chi-squared Automatic Interaction Detection (CHAID) decision tree model was further developed and validated using 10-fold cross-validation, yielding a cross-validation risk value of 0.003, which was comparable to the resubstitution risk value (0.002), indicating good generalization ability and low risk of overfitting. A novel five-dimensional overlapping analysis identified RFW as the core evaluation indicator, with RDW, TFW and R/S as key auxiliary indicators. Conclusion This study delivers a standardized, reproducible technical framework for large-scale screening of saline-alkali-tolerant soybean germplasms. It facilitates global saline-alkali land utilization, accelerates worldwide soybean stress-tolerance breeding, and provides a transferable paradigm for stress tolerance evaluation in other major crops.