Bacteriophages, often referred to as the dark matter of the biosphere, play a critical role in regulating microbial ecosystems and in antibiotic alternatives. Thus, accurate interpretation of their genomes holds significant scientific and practical value. While general-purpose Large Language Models (LLMs) excel at understanding biological texts, their ability to directly interpret raw nucleotide sequences and perform biological reasoning remains underexplored. To address this, we introduce PhageBench, the first benchmark designed to evaluate phage genome understanding by mirroring the workflow of bioinformatics experts. The dataset contains 5,600 high-quality samples covering five core tasks across three stages: Screening, Quality Control, and Phenotype Annotation. Our evaluation of eight LLMs reveals that general-purpose reasoning models significantly outperform random baselines in phage contig identification and host prediction, demonstrating promising potential for genomic understanding. However, they exhibit significant limitations in complex reasoning tasks involving long-range dependencies and fine-grained functional localization. These findings highlight the necessity of developing next-generation models with enhanced reasoning capabilities for biological sequences.
Figures & tables
Figure 1: PhageBench challenges general-purpose LLMs to perform five phage analysis tasks, processing raw nucleotide sequence inputs to generate reasoning-based phenotypic predictions.
Figure 2: The overview of data construction of PhageBench. We filter raw sequences from public repositories, apply task-specific processing rules, and format the resulting samples into standardized instructions for evaluation. For detailed procedures for each task, please refer to Appendix B.1 .
Figure 3: The compositional structure of PhageBench. The sizes of the constructed instruction datasets are labeled below the horizontal lines, and the percentages on the pie charts represent the proportion of data within each major category.
Figure 4: Statistical characteristics of the genomic sequences within the PhageBench dataset. The left panel displays the distribution of sequence sources. The central panel illustrates the probability density of contig lengths. The right panel depicts the distribution of GC content.
Model
T1: Phage Identification
T2: Contamination
T3: Completeness
T4: Lifestyle
T5: Host Prediction
Avg.
Non-reasoning
GPT-4o-mini
50.00
50.08
24.50
48.40
27.75
40.15
LLaMA 4
53.25
49.58
27.10
54.50
28.58
42.60
Qwen3-235b
51.58
49.33
25.40
52.00
28.58
41.38
Reasoning
GPT-OSS-120b
50.08
46.58
24.20
49.50
41.00
42.27
Table 1: Model performance accuracy (%) on PhageBench tasks under zero shot CoT setting. Bold: the best performance. Underlined: the second performance. Avg.: the average across all tasks. T: task.
Figure 5: Performance heatmaps across sequence length bins for Tasks 1–4. Each cell represents the accuracy of a specific model on sequences within a particular length range.
Model
T1: Phage Identification
T2: Contamination
T3: Completeness
T4: Lifestyle
T5: Host Prediction
Average
0-shot
Δ
0-shot
Δ
0-shot
Δ
0-shot
Δ
0-shot
Δ
0-shot
Δ
GPT-4o-mini
50.00
0.00
50.00
+0.08
27.90
− 3.40
48.20
+0.20
26.75
+1.00
40.57
− 0.42
LLaMA-4
51.75
+1.50
51.33
− 1.75
25.60
+1.50
50.60
+3.90
27.83
+0.75
41.42
+1.18
Qwen3-235b
50.00
+1.58
50.42
− 1.09
25.00
+0.40
50.60
+1.40
28.42
+0.16
40.89
+0.49
GPT-5.2
68.50
+2.25
51.42
+2.75
36.60
+6.50
50.50
+1.00
29.17
+6.08
47.24
+3.71
Gemini-3-flash
70.25
+0.58
56.42
+2.41
30.20
+1.90
58.20
− 0.80
64.58
− 2.08
55.93
+0.40
Table 2: Comparison of model accuracy (%) between zero-shot and zero-shot with CoT modes. Grey cells indicate the Δ = AccCoT−Acc0−shot , which is the improvement of CoT. Positive values indicate CoT improvement. Bold values denote the best 0-shot performance per task.
Figure 6: A case study of hallucinated reasoning in Lifestyle Classification. (A) While the model correctly identifies integrase as a key marker, it incorrectly asserts its absence in the sequence.(B) The ground truth annotation confirms the presence of integrase.
Appendix figures & tables12 assets
Supplementary material from the paper’s appendix.
Appendix
Task
Question Template
Phage Contig Identification
Is the following nucleotide sequence derived from a bacteriophage genome?
Contamination Classification
This sequence is a phage-associated contig. Does it contain any DNA originating from a non-phage host genome?
Completeness Classification
What is the estimated genome completeness category of the following bacteriophage sequence?
Lifestyle Classification
What is the most likely lifestyle of the bacteriophage represented by the following genome sequence?
Host Prediction
Which host family/genus/order is the following bacteriophage most likely to infect?
Appendix
Table 3: The specific instructions designed for each task in PhageBench.
Figure 7: Detailed class and subclass distributions across the task 1–4. The pie charts demonstrate the strict balancing strategy applied to primary classes and quality tiers ensuring statistical fairness.
Figure 8: Detailed class distributions of task 5. The pie charts demonstrate the strict balancing strategy applied to fine-grained taxonomic categories.
Figure 9: Distribution of correct answer keys across all tasks. The bar charts verify that the correct options (A, B, C, D) are uniformly distributed, confirming the effectiveness of the option shuffling strategy in mitigating position bias.
Figure 10: The zero shot with CoT prompt.
Figure 11: The zero shot prompt.
Figure 12: Detailed performance analysis of LLMs on PhageBench tasks. (Top) Phage Identification results separated by binary classes and fine-grained biological sources, followed by Contamination Detection across varying contamination ratios; (Bottom) Completeness Estimation across quality tiers, Lifestyle Classification phenotypes, and Host Prediction at three taxonomic resolutions.
Figure 13: Case study of successful Phage Contig Identification.
Figure 14: Case study of failure in Contamination Detection.
Figure 15: Case study of structural hallucination in Completeness Estimation.
Figure 16: Case study of failure in Lifestyle Classification.
Figure 17: Case study of successful Host Prediction.
Large Language Models (LLMs) are increasingly adopted as conversational assistants in genomics, where they are mainly used to reason over biological knowledge, annotations, and analysis outputs through natural language interfaces. However, existing benchmarks either focus on specialized DNA models trained for sequence prediction or evaluate biological knowledge using text-only questions, leaving the behavior of general-purpose LLMs when directly exposed to raw genome sequences underexplored. We introduce GenomeQA, a benchmark designed to provide a controlled evaluation setting for general-purpose LLMs on sequence-based genome inference tasks. GenomeQA comprises 5,200 samples drawn from multiple biological databases, with sequence lengths ranging from 6 to 1,000 base pairs (bp), spanning six task families: Enhancer and Promoter Identification, Splice Site Identification, Taxonomic Classification, Histone Mark Prediction, Transcription Factor Binding Site Prediction, and TF Motif Prediction. Across six frontier LLMs, we find that models consistently outperform random baselines and can exploit local sequence signals such as GC content and short motifs, while performance degrades on tasks that require more indirect or multi-step inference over sequence patterns. GenomeQA establishes a diagnostic benchmark for studying and improving the use of general-purpose LLMs on raw genomic sequences.
Weicai Long, Yusen Hou, Junning Feng +4
Hong Kong University of Science and Technology (Guangzhou) · The University of Hong Kong · Peking University
Nucleotide sequences constitute the fundamental genetic basis of biological systems, rendering viral genomic analysis critical for biomedical advancement. Despite progress in biological foundation models, specifically nucleotide foundation models (NFMs), the field lacks a unified standard for viral genomics to facilitate community development and enforce biosecurity constraints. To address this, we introduce ViroBench, the first comprehensive and large-scale benchmark specifically designed for NFMs in viral settings. ViroBench evaluates models across two critical dimensions: biological understanding and latent biosecurity risk, covering 18 diverse scenarios within 4 task types. Extensive evaluation of 66 NFMs across diverse architectures yields three critical conclusions. Firstly, NFMs exhibit a performance degradation in biological understanding under phylogenetic and temporal shifts, indicating weak extrapolation capabilities. Secondly, generation tasks reveal a decoupling between statistical likelihood and biological functional validity, posing latent biosecurity risks. Thirdly, controlled ablation studies reveal that taxonomic diversity in pretraining data outweighs parameter scale. Specifically, a lightweight baseline trained on diverse data achieves a 67.5% performance gain over its original model. Overall, ViroBench provides interpretable, diagnostic evaluations and a reproducible measurement framework for future research on viral nucleotide foundation models. The datasets and code are publicly available at https://github.com/QIANJINYDX/ViroBench.
Dongxin Ye, Fang Hu, Han Hu +6
Shanghai Innovation Institute Shanghai, China · University of Electronic Science and Technology of China Chengdu, China · Fudan University Shanghai, China +7
Multi-omics sequences contain complex biological patterns, yet deciphering their mechanisms for automated scientific discovery remains challenging. As large language models (LLMs) interpret these sequences, evaluating both predictions and scientific reasoning is critical. However, existing benchmarks for multi-omics sequence tasks rely on classification and regression metrics, neglecting whether models grasp the underlying biological evidence. We introduce OmicsBench, the first reasoning benchmark for multi-omics sequences, comprising 1,160 expert-validated questions across six tasks spanning DNA regulation, RNA processing, and protein function. OmicsBench requires traceable evidence chains, evaluated using instance-specific rubrics developed with domain experts. Evaluating 17 LLMs reveals an inverse relationship: while scientific LLMs outperform general-purpose LLMs in sequence classification accuracy, they fail to provide valid evidence to support their predictions. One plausible interpretation is shortcut learning: specialized models may rely on statistical patterns rather than the biological mechanisms needed for scientific discovery. Motivated by this finding, we introduce tool-augmented on-policy distillation (TA-OPD), a post-training method to align sequence prediction with evidence-grounded biological reasoning. Across five Qwen3.5 models spanning 0.8B to 27B parameters, TA-OPD consistently strengthens biological evidence grounding while improving predictive performance on most tasks. These gains persist across model scales, indicating that stronger sequence reasoning does not arise solely from increased model capacity, but can be improved through evidence-aware training. Together, OmicsBench and TA-OPD provide a framework for diagnosing reasoning failures in multi-omics LLMs and a path toward models whose predictions are better grounded in biologically meaningful evidence.
Jie Ying, Zhefan Wang, Zihong Chen +11
Shanghai Artificial Intelligence Laboratory · Yazhouwan National Laboratory · Shanghai Innovation Institute +2